CN106248657B - One kind is based on dendroid Fe2O3The preparation method and application of@Au Electrochemiluminescsensor sensor - Google Patents
One kind is based on dendroid Fe2O3The preparation method and application of@Au Electrochemiluminescsensor sensor Download PDFInfo
- Publication number
- CN106248657B CN106248657B CN201610803682.9A CN201610803682A CN106248657B CN 106248657 B CN106248657 B CN 106248657B CN 201610803682 A CN201610803682 A CN 201610803682A CN 106248657 B CN106248657 B CN 106248657B
- Authority
- CN
- China
- Prior art keywords
- solution
- concentration
- under
- ultra
- dendroid
- 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
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
-
- 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
Abstract
Dendroid Fe is based on the present invention relates to one kind2O3The preparation method and application of@Au Electrochemiluminescsensor sensor, belongs to electrochemical luminous sensor field, using tris (bipyridine) ruthenium as electrochemical luminescence signals source, utilizes dendroid Fe2O3Bio-compatibility excellent@Au and big specific surface area increase the supported quantity of antibody, use Pd graphene/Fe3O4‑Ru(bpy)3 2+As secondary antibody label, according to the difference of electrochemical luminescence signals intensity, the detection to liver cancer marker is realized.
Description
Technical field
Dendroid Fe is based on the present invention relates to one kind2O3The preparation method and application of@Au Electrochemiluminescsensor sensor,
Specifically related to a kind of tris (bipyridine) ruthenium utilizes dendroid Fe as luminescent material2O3Bio-compatibility excellent@Au and big ratio
Surface area increases the supported quantity of antibody as base material, uses Pd-graphene/Fe3O4-Ru(bpy)3 2+Marked as secondary antibody
Thing, belongs to electrochemical luminescence detection technique field.
Background technology
In recent years, the incidence of disease of liver cancer remains high, and the death rate increases year by year, and reason is discovery to liver cancer not mostly
It is enough timely, optimal treatment time is missed to while finding to be later period of hepatocarcinoma.Found by research in recent years, alpha-fetoprotein
AFP, squamous cell carcinoma antigen SCCA, CEA, carbohydrate antigen CA15-3 etc. can as liver cancer marker, they
Played an important role in the early prevention of liver cancer and treatment.Concentration progress to liver cancer marker in serum is quick, sensitive
Detection has very important clinical value and meaning, and the death rate of liver cancer can be reduced to the full extent, and extension liver cancer is suffered from
In the existence time limit of person, be that patient strives for more therapy apparatus meetings.
Method currently used for detection liver cancer marker is mainly radioimmunoassays and Chemiluminescence immunoassay, but these
Method exist can not immediately quick detection, with the drawback such as radioactive pollution and complex operation, and electrochemical immunosensor
These drawbacks can be overcome well, and it is a kind of immunosensor for being recognized and being built based on antigen and antibody specificity, with
By its preparation is simple, sensitivity is high, test limit is low, fast response time advantage, the inspection of rapid sensitive can be carried out to antigen
Survey.
The content of the invention
The purpose of the present invention is that there is provided one kind is simple and quick the problem of presence for existing liver cancer marker detection method
Reliably it is based on dendroid Fe2O3@Au and Pd-graphene/Fe3O4-Ru(bpy)3 2+Electrochemical luminescence sensor preparation
Methods and applications, are realized to the quick, sensitive, special of liver cancer marker, efficient detection.
Technical scheme is as follows：
1. one kind is based on dendroid Fe2O3The preparation method of@Au Electrochemiluminescsensor sensor, it is characterised in that bag
Include following steps：
（1）The mm of diameter 4 glass-carbon electrode is taken turns doing with 1.0 μm, 0.3 μm, 0.05 μm of aluminum oxide polishing powder successively
Polishing, it is clean with ultrapure water；
（2）The μ L of drop coating 6, concentration are 2 ~ 4 mg/mL antibody A b1Capture base material Fe2O3@Au/Ab1Solution is to electricity
Pole surface, 4 °C are dried；
（3）The bovine serum albumin solution that 3 μ L, mass fraction are 1 ~ 3% is added dropwise, with the non-specific of enclosed-electrode surface
Property avtive spot, rinse electrode surface with pH 7.4 phosphate buffer solution solution, 4 °C are dried；
（4）0.5 ~ 2 h of hatching under 6 μ L, certain density determinand antigen, 4 °C is added dropwise, with pH 7.4 phosphate
Cushioning liquid solution rinses electrode surface, and 4 °C are dried；
（5）The μ L of drop coating 6, concentration are 2 ~ 4 mg/mL secondary antibody Ab2Label Pd-graphene/Fe3O4 -Ru
(bpy)3 2+/Ab2Solution, electrode surface is rinsed with pH 7.4 phosphate buffer solution solution, and after 4 °C are dried, one kind is made
Based on dendroid Fe2O3@Au Electrochemiluminescsensor sensor.
2. antibody capture base material Fe2O3@Au/Ab1The preparation of solution
（1）0.05 ~ 0.20 g potassium ferricyanide solid is dissolved in 5 ~ 50 mL ultra-pure waters, with 0.001 ~ 0.020
Mol/L sodium hydroxide solution regulation pH to 10.0 ~ 12.0, continues to stir 1 ~ 15 min, mixed solution is put into high pressure
In reactor, 6 ~ 24 h are reacted under 60 ~ 150 °C, product is centrifuged under 6000 ~ 8000 rpm and nothing is used
Water-ethanol and ultra-pure water are washed three times respectively, are put under vacuum drying chamber, 30 ~ 60 °C and are dried 2 ~ 5 h, are made brick-red
Dendroid Fe2O3；
（2）40 ~ 100 mL, mass fraction are boiled for 0.001 ~ 0.010% chlorauric acid solution, 1 ~ 3 is added
ML, mass fraction are 0.1 ~ 1.0% trisodium citrate aqueous solution, are heated to reflux 5 ~ 20 min, treat that solution colour becomes wine
Red, room temperature is cooled to by solution, is made under solution of gold nanoparticles, 4 °C and is kept in dark place；
（3）By 0.5 ~ 3 mg dendroid Fe2O3It is distributed in 1 mL ultra-pure waters, adds 0.5 ~ 1.5 mL gold nanos
Particle solution, mixed solution is vibrated at room temperature 12 ~ 32 h, is centrifuged, and removes excessive golden nanometer particle, is made
Fe2O3@Au, by product Fe2O3@Au are re-dispersed into 1 mL ultra-pure waters, and base material Fe is made2O3@Au solution；
（4）It is 2 ~ 4 mg/mL Fe in 1 ~ 3 mL, concentration2O3In@Au solution, adding 100 ~ 400 μ L, concentration is
10 μ g/mL determinand antibody A b1Solution, 24 h of vibration hatching, are centrifuged off excessive determinand antibody A b under 4 °C1,
In the phosphate buffer solution solution that product is distributed to 1 mL, pH 7.4, antibody capture base material Fe is made2O3@Au/Ab1
Solution, is stored in standby under 4 °C.
3. secondary antibody label Pd-graphene/Fe3O4-Ru(bpy)3 2+/Ab2The preparation of solution
（1）It is that 12 mol/L hydrochloric acid solutions are added in 5 ~ 30 mL ultra-pure water by 0.1 ~ 2 mL, concentration, then
0.5 ~ 6.0 g ferric trichlorides solid and 0.5 ~ 6.0 g frerrous chloride solids are added into solution, under continuous stirring, will
Mixed solution is added dropwise in the sodium hydroxide solution that 100 ~ 500 mL, concentration are 0.5 ~ 5.0 mol/L, 4000
Be centrifuged under ~ 8000 rpm rotating speed, with milli-Q water three times, backward precipitation in add 200 ~ 600 mL,
Concentration is 0.01 ~ 0.04 mol/L hydrochloric acid solution, and 5 ~ 15min is centrifuged under 6000 ~ 9000 rpm rotating speed,
Abandoning supernatant, adds ultra-pure water and carries out the Fe that ultrasound obtains yellow3O4Nano sol；
（2）By 1 ~ 5 mL Fe3O4Nano sol is added to 40 ~ 200 mL, concentration and aoxidized for 1.0 ~ 3.0 mg/mL
In graphene dispersion suspension, under agitation, 0.10 ~ 0.60 g sodium hydrogensulfite solids are added, reacted under 80 ~ 160 °C
2 ~ 6 h, are dialysed one week after cooling with ultra-pure water, it is freeze-dried after can obtain the graphene/Fe of 3D structures3O4；
（3）0.06 ~ 0.20 g tetrachloro-palladium acid sodium solids and 0.06 ~ 0.20 g sodium citrate solids are dissolved in 10 ~
In 100 mL ultra-pure waters, it is that 0.005 ~ 0.020 mol/L sodium borohydrides are molten that 0.1 ~ 4.0 mL, concentration are added into solution
Liquid, persistently stirs 30 ~ 90 s, and the Pd nano-particle solutions of black are made；
（4）By 1 ~ 3 mL, the graphene/Fe that concentration is 2 mg/mL3O4The Pd nano-particles of solution and 1 ~ 3 mL
Solution mix, lucifuge vibrate 24 ~ 48 h, centrifuge abandoning supernatant, be distributed in 1 mL ultra-pure waters, with 1 ~ 3 mL,
Concentration is 1 × 10-3 mol/L Ru(bpy)3 2+Solution mixing 24 ~ 48 h of vibration, centrifuge abandoning supernatant, are distributed to 1
In mL ultra-pure waters, Pd-graphene/Fe is made3O4-Ru(bpy)3 2+Solution；
（5）It is 2 ~ 4 mg/mL Pd-graphene/Fe in 1 ~ 3 mL, concentration3O4-Ru(bpy)3 2+In solution, add
100 ~ 400 μ L, concentration are 10 μ g/mL determinand antibody A b2Solution, 24 h of vibration hatching, are centrifuged off under 4 °C
Excessive determinand antibody A b2, in the phosphate buffer solution solution that product is distributed to 1 mL, pH 7.4, secondary antibody mark is made
Remember thing Pd-graphene/Fe3O4-Ru(bpy)3 2+/Ab2Solution, is stored in standby under 4 °C.
4. the electrochemical luminescence sensor prepared is used for the Electrochemical Detection of testing sample
（1）Tested using the three-electrode system of electrochemical workstation, Ag/AgCl electrodes are used as reference electrode, platinum filament
Electrode is that prepared electrochemical luminescence sensor is working electrode, by electrochemical workstation and chemiluminescence detection to electrode
Instrument links together the high pressure of photomultiplier being set to 800 V, and cyclic voltammetry scan potential range is 0 ~ 1.2 V, is swept
Speed is retouched for 0.1 V/s；
（2）In 10 mL, pH 6.4 ~ 8.4 containing the phosphate buffer solution that concentration is 35 ~ 65 mmol/L triethylamines
In solution, by electrochemical luminescence system, the electrochemical luminescence signals intensity produced to the determinand antigen of various concentrations is detected,
Drawing curve；
（3）Testing sample solution is detected instead of determinand antigen.
5. determinand antigen is selected from one of following liver cancer marker：AFP, squamous cell carcinoma antigen SCCA, cancer
Embryonal antigen CEA, carbohydrate antigen CA15-3.
The useful achievement of the present invention
（1）Electrochemical luminescence sensor preparation method, with Ru (bpy)3 2+For luminescent material, utilize Ru (bpy)3 2+Good
Optical property, the sensor of structure has higher sensitivity.
（2）With Fe2O3@Au are used as antibody capture substrate, Pd-graphene/Fe3O4-Ru(bpy)3 2+Marked as secondary antibody
Thing, Fe2O3@Au and Pd-graphene/Fe3O4-Ru(bpy)3 2+Bio-compatibility is good, and the advantages of specific surface area is big effectively increases
The supported quantity of antibody is added.
（3）Electrochemical luminescence sensor prepared by the present invention is used for the detection of liver cancer marker, simple to operate, and reaction is fast
Speed, signal response range is wide, it is possible to achieve simple, quick, sensitive, specific detection.
Embodiment 1 is a kind of to be based on dendroid Fe2O3The preparation method of@Au Electrochemiluminescsensor sensor
（1）The mm of diameter 4 glass-carbon electrode is taken turns doing with 1.0 μm, 0.3 μm, 0.05 μm of aluminum oxide polishing powder successively
Polishing, it is clean with ultrapure water；
（2）6 μ L, 2 mg/mL antibody A b is added dropwise1Capture base material Fe2O3@Au/Ab1Solution is to electrode surface, 4 °
C dries；
（3）The bovine serum albumin solution that 3 μ L, mass fraction are 1% is added dropwise, it is living with the non-specificity on enclosed-electrode surface
Property site, rinse electrode surface with pH 7.4 phosphate buffer solution solution, 4 °C are dried；
（4）It is added dropwise under 6 μ L, certain density determinand antigen, 4 °C and hatches 0.5 h, with pH 7.4 phosphate-buffered
Solutions Solution rinses electrode surface, and 4 °C are dried；
（5）6 μ L, concentration is added dropwise for 2 mg/mL secondary antibodies Ab2Label Pd-graphene/Fe3O4-Ru(bpy)3 2+ /Ab2
Solution, electrode surface is rinsed with pH 7.4 phosphate buffer solution solution, after 4 °C are dried, and is made a kind of and is based on dendroid
Fe2O3@Au Electrochemiluminescsensor sensor.
Embodiment 2 is a kind of to be based on dendroid Fe2O3The preparation method of@Au Electrochemiluminescsensor sensor
（1）The mm of diameter 4 glass-carbon electrode is taken turns doing with 1.0 μm, 0.3 μm, 0.05 μm of aluminum oxide polishing powder successively
Polishing, it is clean with ultrapure water；
（2）6 μ L, 3 mg/mL antibody capture base material Fe is added dropwise2O3@Au/Ab1Solution and electrode surface, 4 °C dry in the air
It is dry；
（3）The bovine serum albumin solution that 3 μ L, mass fraction are 2% is added dropwise, it is living with the non-specificity on enclosed-electrode surface
Property site, rinse electrode surface with pH 7.4 phosphate buffer solution solution, 4 °C are dried；
（4）It is added dropwise under 6 μ L, certain density determinand antigen, 4 °C and hatches 1.5 h, with pH 7.4 phosphate-buffered
Solutions Solution rinses electrode surface, and 4 °C are dried；
（5）6 μ L, concentration is added dropwise for 3 mg/mL secondary antibodies Ab2Label Pd-graphene/Fe3O4-Ru(bpy)3 2+ /Ab2
Solution, electrode surface is rinsed with pH 7.4 phosphate buffer solution solution, after 4 °C are dried, and is made a kind of and is based on dendroid
Fe2O3@Au Electrochemiluminescsensor sensor.
Embodiment 3 is a kind of to be based on dendroid Fe2O3The preparation method of@Au Electrochemiluminescsensor sensor
（1）The mm of diameter 4 glass-carbon electrode is taken turns doing with 1.0 μm, 0.3 μm, 0.05 μm of aluminum oxide polishing powder successively
Polishing, it is clean with ultrapure water；
（2）6 μ L, 4 mg/mL antibody A b is added dropwise1Capture base material Fe2O3@Au/Ab1Solution and electrode surface, 4 °C
Dry；
（3）The bovine serum albumin solution that 3 μ L, mass fraction are 3% is added dropwise, it is living with the non-specificity on enclosed-electrode surface
Property site, rinse electrode surface with pH 7.4 phosphate buffer solution solution, 4 °C are dried；
（4）It is added dropwise under 6 μ L, certain density determinand antigen, 4 °C and hatches 2 h, the phosphate-buffered with pH 7.4 is molten
Liquor rinses electrode surface, and 4 °C are dried；
（5）6 μ L, concentration is added dropwise for 4 mg/mL secondary antibodies Ab2Label Pd-graphene/Fe3O4-Ru(bpy)3 2+ /Ab2
Solution, electrode surface is rinsed with pH 7.4 phosphate buffer solution solution, after 4 °C are dried, and is made a kind of and is based on dendroid
Fe2O3@Au Electrochemiluminescsensor sensor.
The antibody capture base material Fe of embodiment 42O3@Au/Ab1The preparation of solution
（1）0.05 g potassium ferricyanide solid is dissolved in 10 mL ultra-pure waters, the sodium hydroxide with 0.001 mol/L is molten
Liquid adjusts pH to 10.0, continues to stir 5 min, mixed solution is put into autoclave, 8 h are reacted under 80 °C, will be produced
Product are centrifuged and washed respectively three times with absolute ethyl alcohol and ultra-pure water under 6000 rpm, are put under vacuum drying chamber, 30 °C
2 h are dried, brick-red dendroid Fe is made2O3；
（2）40 mL, mass fraction are boiled for 0.001% chlorauric acid solution, it is 0.1 % to add 1 mL, mass fraction
Trisodium citrate aqueous solution, be heated to reflux 5 min, treat that solution colour becomes claret, solution is cooled to room temperature, be made gold
Nano-particle solution, is kept in dark place under 4 °C；
（3）By 0.5 mg dendroid Fe2O3It is distributed in 1 mL ultra-pure waters, adds 0.5 mL solution of gold nanoparticles, will
Mixed solution vibrates 12 h at room temperature, centrifuges, and removes excessive golden nanometer particle, and Fe is made2O3@Au, by product
Fe2O3@Au are re-dispersed into 1 mL ultra-pure waters, and base material Fe is made2O3@Au solution；
（4）It is 2 mg/mL Fe in 1 mL, concentration2O3In@Au solution, it is the to be measured of 10 μ g/mL to add 100 μ L, concentration
Thing antibody A b1Solution, 24 h of vibration hatching, are centrifuged off excessive determinand antibody A b under 4 °C1, product is distributed to 1
In mL, pH 7.4 phosphate buffer solution solution, antibody capture base material Fe is made2O3@Au/Ab1Solution, is stored in 4 °
It is standby under C.
The antibody capture base material Fe of embodiment 52O3@Au/Ab1The preparation of solution
（1）0.15 g potassium ferricyanide solid is dissolved in 25 mL ultra-pure waters, with 0.01 mol/L sodium hydroxide solution
PH to 11.0 is adjusted, continues to stir 8 min, mixed solution is put into autoclave, 12 h are reacted under 120 °C, will be produced
Product are centrifuged and washed respectively three times with absolute ethyl alcohol and ultra-pure water under 7000 rpm, are put under vacuum drying chamber, 40 °C
3 h are dried, brick-red dendroid Fe is made2O3；
（2）80 mL, mass fraction are boiled for 0.005% chlorauric acid solution, it is 0.5% to add 1.5 mL, mass fraction
Trisodium citrate aqueous solution, be heated to reflux 10 min, treat that solution colour becomes claret, solution is cooled to room temperature, be made
Solution of gold nanoparticles, is kept in dark place under 4 °C；
（3）By 1 mg dendroid Fe2O3It is distributed in 1 mL ultra-pure waters, adds 1 mL solution of gold nanoparticles, will mix
Solution vibrates 18 h at room temperature, centrifuges, and removes excessive golden nanometer particle, and Fe is made2O3@Au, by product Fe2O3@Au
It is re-dispersed into 1 mL ultra-pure waters, base material Fe is made2O3@Au solution；
（4）It is 3 mg/mL Fe in 2 mL, concentration2O3In@Au solution, it is the to be measured of 10 μ g/mL to add 200 μ L, concentration
Thing antibody A b1Solution, 24 h of vibration hatching, are centrifuged off excessive determinand antibody A b under 4 °C1, product is distributed to 1
In mL, pH 7.4 phosphate buffer solution solution, antibody capture base material Fe is made2O3@Au/Ab1Solution, is stored in 4 °
It is standby under C.
The antibody capture base material Fe of embodiment 62O3@Au/Ab1The preparation of solution
（1）0.19 g potassium ferricyanide solid is dissolved in 40 mL ultra-pure waters, the sodium hydroxide with 0.015 mol/L is molten
Liquid adjusts pH to 12.0, continues to stir 15 min, mixed solution is put into autoclave, 24 h are reacted under 140 °C,
Product is centrifuged and washed respectively three times with absolute ethyl alcohol and ultra-pure water under 8000 rpm, vacuum drying chamber, 50 is put into
4 h are dried under °C, brick-red dendroid Fe is made2O3；
（2）100 mL, mass fraction are boiled for 0.010% chlorauric acid solution, it is 1.0% to add 2 mL, mass fraction
Trisodium citrate aqueous solution, be heated to reflux 15 min, treat that solution colour becomes claret, solution is cooled to room temperature, be made
Solution of gold nanoparticles, is kept in dark place under 4 °C；
（3）By 2 mg dendroid Fe2O3It is distributed in 1 mL ultra-pure waters, adds 1.5 mL solution of gold nanoparticles, will be mixed
Close solution and vibrate 32 h at room temperature, centrifuge, remove excessive golden nanometer particle, Fe is made2O3@Au, by product Fe2O3@
Au is re-dispersed into 1 mL ultra-pure waters, and base material Fe is made2O3@Au solution；
（4）It is 4 mg/mL Fe in 3 mL, concentration2O3In@Au solution, it is the to be measured of 10 μ g/mL to add 400 μ L, concentration
Thing antibody A b1Solution, 24 h of vibration hatching, are centrifuged off excessive determinand antibody A b under 4 °C1, product is distributed to 1
In mL, pH 7.4 phosphate buffer solution solution, antibody capture base material Fe is made2O3@Au/Ab1Solution, is stored in 4 °
It is standby under C.
The secondary antibody label Pd-graphene/Fe of embodiment 73O4-Ru(bpy)3 2+/Ab2The preparation of solution
（1）It is that 12 mol/L hydrochloric acid solutions are added in 13 mL ultra-pure water by 0.4 mL, concentration, then in solution
2.6 g ferric trichlorides solids and 1.6g frerrous chloride solids are added, under continuous stirring, mixed solution is added dropwise to
125 mL, concentration is in 1.5 mol/L sodium hydroxide solution, are centrifuged under 4000 rpm rotating speed, with ultrapure
Water washing three times, backward precipitation in add the hydrochloric acid solution that 250 mL, concentration are 0.01 mol/L, in 6000 rpm rotating speed
5 min of lower centrifugation, abandoning supernatant adds ultra-pure water and carries out the Fe that ultrasound obtains yellow3O4Nano sol；
（2）By 2.5 mL Fe3O4Nano sol is added to 50 mL, concentration for 1.5 mg/mL graphene oxide dispersible suspensions
In, under agitation, 0.22 g sodium hydrogensulfite solids are added, 3 h are reacted under 95 °C, dialysed one week with ultra-pure water after cooling,
Graphene/Fe is obtained after freeze-dried3O4；
（3）0.07 g tetrachloro-palladium acid sodium solids and 0.07 g sodium citrate solids are dissolved in 20 mL ultra-pure waters, to solution
0.6 mL of middle addition, concentration is 0.01 mol/L sodium borohydride solutions, persistently stirs 30 s, and the Pd nano-particles that black is made are molten
Liquid；
（4）By 1 mL, the graphene/Fe that concentration is 2 mg/mL3O4Solution is mixed with 1 mL Pd nano-particle solutions,
Lucifuge vibrates 24 h, centrifuges abandoning supernatant, is distributed in 1 mL ultra-pure waters, is 1 × 10 with 1 mL, concentration-3 mol/L
Ru(bpy)3 2+Solution mixing vibration 24h, centrifuges abandoning supernatant, is distributed in 1 mL ultra-pure waters, Pd- is made
graphene/Fe3O4-Ru(bpy)3 2+Solution；
（5）It is 2 mg/mL Pd-graphene/Fe in 1 mL, concentration3O4-Ru(bpy)3 2+In solution, 100 μ L of addition,
Concentration is 10 μ g/mL determinand antibody A b2Solution, 24 h of vibration hatching under 4 °C, is centrifuged off excessive determinand and resists
Body Ab2, in the phosphate buffer solution solution that product is distributed to 1 mL, pH 7.4, secondary antibody label Pd- is made
graphene/Fe3O4-Ru(bpy)3 2+/Ab2Solution, is stored in standby under 4 °C.
The secondary antibody label Pd-graphene/Fe of embodiment 83O4-Ru(bpy)3 2+/Ab2The preparation of solution
（1）It is that 12 mol/L hydrochloric acid solutions are added in 20 mL ultra-pure water by 1.5 mL, concentration, then in solution
3.0 g ferric trichlorides solids and 4.0 g frerrous chloride solids are added, under continuous stirring, mixed solution is added dropwise to
300 mL, concentration is in 3.0 mol/L sodium hydroxide solution, are centrifuged under 6000 rpm rotating speed, with ultrapure
Water washing three times, backward precipitation in add the hydrochloric acid solution that 400 mL, concentration are 0.03 mol/L, in 7000 rpm rotating speed
10 min of lower centrifugation, abandoning supernatant adds ultra-pure water and carries out the Fe that ultrasound obtains yellow3O4Nano sol；
（2）By 2 mL Fe3O4Nano sol is added to 100 mL, concentration for 2.0 mg/mL graphene oxide dispersible suspensions
In, under agitation, 0.40 g sodium hydrogensulfite solids are added, 4 h are reacted under 120 °C, with ultra-pure water dialysis one after cooling
Week, it is freeze-dried after obtain graphene/Fe3O4；
（3）0.15 g tetrachloro-palladium acid sodium solids and 0.15 g sodium citrate solids are dissolved in 80 mL ultra-pure waters, to solution
3.0 mL of middle addition, concentration are 0.015 mol/L sodium borohydride solutions, persistently stir 60 s, and the Pd nano-particles of black are made
Solution；
（4）By 2 mL, the graphene/Fe that concentration is 2 mg/mL3O4Solution is mixed with 2 mL Pd nano-particle solutions,
Lucifuge vibrates 32 h, centrifuges abandoning supernatant, is distributed in 1 mL ultra-pure waters, is 1 × 10 with 2 mL, concentration-3 mol/L
Ru(bpy)3 2+Solution mixing 32 h of vibration, centrifuge abandoning supernatant, are distributed in 1 mL ultra-pure waters, Pd- is made
graphene/Fe3O4-Ru(bpy)3 2+Solution；
（5）It is 3 mg/mL Pd-graphene/Fe in 2 mL, concentration3O4-Ru(bpy)3 2+In solution, 200 μ L of addition,
Concentration is 10 μ g/mL determinand antibody A b2Solution, 24 h of vibration hatching under 4 °C, is centrifuged off excessive determinand and resists
Body Ab2, in the phosphate buffer solution solution that product is distributed to 1 mL, pH 7.4, secondary antibody label Pd- is made
graphene/Fe3O4-Ru(bpy)3 2+/Ab2Solution, is stored in standby under 4 °C.
The secondary antibody label Pd-graphene/Fe of embodiment 93O4-Ru(bpy)3 2+/Ab2The preparation of solution
（1）It is that 12 mol/L hydrochloric acid solutions are added in 30 mL ultra-pure water by 2 mL, concentration, adds then in solution
Enter 6.0 g ferric trichlorides solids and 6.0 g frerrous chloride solids, under continuous stirring, mixed solution is added dropwise to
500 mL, concentration is in 5.0 mol/L sodium hydroxide solution, are centrifuged under 8000 rpm rotating speed, with ultrapure
Water washing three times, backward precipitation in add the hydrochloric acid solution that 600 mL, concentration are 0.04 mol/L, in 9000 rpm rotating speed
15 min of lower centrifugation, abandoning supernatant adds ultra-pure water and carries out the Fe that ultrasound obtains yellow3O4Nano sol；
（2）By 5 mL Fe3O4Nano sol is added to 200 mL, concentration for 3.0 mg/mL graphene oxide dispersible suspensions
In, under agitation, 0.60 g sodium hydrogensulfite solids are added, 6 h are reacted under 160 °C, with ultra-pure water dialysis one after cooling
Week, it is freeze-dried after obtain graphene/Fe3O4；
（3）0.20 g tetrachloro-palladium acid sodium solids and 0.20 g sodium citrate solids are dissolved in 100 mL ultra-pure waters, Xiang Rong
4.0 mL are added in liquid, concentration is 0.020 mol/L sodium borohydride solutions, persistently stirs 90 s, and the Pd nanoparticles of black are made
Sub- solution；
（4）By 3 mL, the graphene/Fe that concentration is 2 mg/mL3O4Solution is mixed with 3 mL Pd nano-particle solutions,
Lucifuge vibrates 48 h, centrifuges abandoning supernatant, is distributed in 1 mL ultra-pure waters, is 1 × 10 with 3 mL, concentration-3 mol/L
Ru (bpy)3 2+Solution mixing 48 h of vibration, centrifuge abandoning supernatant, are distributed in 1 mL ultra-pure waters, Pd- is made
graphene/Fe3O4-Ru(bpy)3 2+Solution；
（5）It is 4 mg/mL Pd-graphene/Fe in 3 mL, concentration3O4-Ru(bpy)3 2+In solution, 400 μ L of addition,
Concentration is 10 μ g/mL determinand antibody A b2Solution, 24 h of vibration hatching under 4 °C, is centrifuged off excessive determinand and resists
Body Ab2, in the phosphate buffer solution solution that product is distributed to 1 mL, pH 7.4, secondary antibody label Pd- is made
graphene/Fe3O4-Ru(bpy)3 2+/Ab2Solution, is stored in standby under 4 °C.
The detection of the AFP of embodiment 10
（1）Tested using the three-electrode system of electrochemical workstation, Ag/AgCl electrodes are used as reference electrode, platinum filament
Electrode is that prepared electrochemical luminescence sensor is working electrode, by electrochemical workstation and chemiluminescence detection to electrode
Instrument links together the high pressure of photomultiplier being set to 800 V, and cyclic voltammetry scan potential range is 0 ~ 1.2 V, is swept
Speed is retouched for 0.1 V/s；
（2）It is molten for the phosphate buffer solution of 35 ~ 65 mmol/L triethylamines containing concentration in 10 mL, pH 6.4 ~ 8.4
In liquid, by electrochemical luminescence system, detect and the squamous cell carcinoma antigen SCCA of the various concentrations electrochemical luminescences produced are believed
Number intensity, drawing curve, it is the ng/mL of 0.001 pg/mL ~ 1 to measure the range of linearity, and detection is limited to 0.29 fg/mL；
（3）Testing sample solution is detected instead of AFP.
The squamous cell carcinoma antigen SCCA of embodiment 11 detection
（1）Tested using the three-electrode system of electrochemical workstation, Ag/AgCl electrodes are used as reference electrode, platinum filament
Electrode is that prepared electrochemical luminescence sensor is working electrode, by electrochemical workstation and chemiluminescence detection to electrode
Instrument links together the high pressure of photomultiplier being set to 800 V, and cyclic voltammetry scan potential range is 0 ~ 1.2 V, is swept
Speed is retouched for 0.1 V/s；
（2）It is molten for the phosphate buffer solution of 35 ~ 65 mmol/L triethylamines containing concentration in 10 mL, pH 6.4 ~ 8.4
In liquid, by electrochemical luminescence system, detect and the squamous cell carcinoma antigen SCCA of the various concentrations electrochemical luminescences produced are believed
Number intensity, drawing curve, it is the ng/mL of 0.001 pg/mL ~ 1 to measure the range of linearity, and detection is limited to 0.29 fg/mL；
（3）Testing sample solution is detected instead of squamous cell carcinoma antigen SCCA.
The detection of the CEA of embodiment 12
（1）Tested using the three-electrode system of electrochemical workstation, Ag/AgCl electrodes are used as reference electrode, platinum filament
Electrode is that prepared electrochemical luminescence sensor is working electrode, by electrochemical workstation and chemiluminescence detection to electrode
Instrument links together the high pressure of photomultiplier being set to 800 V, and cyclic voltammetry scan potential range is 0 ~ 1.2 V, is swept
Speed is retouched for 0.1 V/s；
（2）In 10 mL, pH 6.4 ~ 8.4 containing the phosphate buffer solution that concentration is 35 ~ 65 mmol/L triethylamines
In solution, by electrochemical luminescence system, detect that the electrochemical luminescence signals produced to the CEA of various concentrations are strong
Degree, drawing curve, it is the ng/mL of 0.001 pg/mL ~ 1 to measure the range of linearity, and detection is limited to 0.29 fg/mL；
（3）Testing sample solution is detected instead of CEA.
Claims (5)
1. one kind is based on dendroid Fe2O3The preparation method of@Au Electrochemiluminescsensor sensor, it is characterised in that including following
Step：
（1）The mm of diameter 4 glass-carbon electrode is taken turns doing into polishing with 1.0 μm, 0.3 μm, 0.05 μm of aluminum oxide polishing powder successively
Processing, it is clean with ultrapure water；
（2）The μ L of drop coating 6, concentration are 2 ~ 4 mg/mL antibody A b1Capture base material Fe2O3@Au/Ab1Solution is to electrode table
Face, 4 °C are dried；
（3）The bovine serum albumin solution that 3 μ L, mass fraction are 1 ~ 3% is added dropwise, it is living with the non-specificity on enclosed-electrode surface
Property site, rinse electrode surface with pH 7.4 phosphate buffer solution, 4 °C are dried；
（4）0.5 ~ 2 h of hatching under 6 μ L, certain density determinand antigen, 4 °C is added dropwise, with pH 7.4 phosphate-buffered
Solutions Solution rinses electrode surface, and 4 °C are dried；
（5）The μ L of drop coating 6, concentration are 2 ~ 4 mg/mL secondary antibody Ab2Label Pd-graphene/Fe3O4- Ru(bpy)3 2+ /
Ab2Solution, rinses electrode surface, 4 °C are dried with pH 7.4 phosphate buffer solution solution, is made a kind of and is based on dendroid
Fe2O3@Au Electrochemiluminescsensor sensor.
2. it is as claimed in claim 1 a kind of based on dendroid Fe2O3The preparation method of@Au Electrochemiluminescsensor sensor, its
It is characterised by, described antibody A b1Capture base material Fe2O3@Au/Ab1Solution preparation step is as follows：
（1）0.05 ~ 0.20 g potassium ferricyanide solid is dissolved in 5 ~ 50 mL ultra-pure waters, with 0.001 ~ 0.020
Mol/L sodium hydroxide solution regulation pH to 10.0 ~ 12.0, continues to stir 1 ~ 15 min, mixed solution is put into high pressure
In reactor, 6 ~ 24 h are reacted under 60 ~ 150 °C, product is centrifuged under 6000 ~ 8000 rpm and nothing is used
Water-ethanol and ultra-pure water are washed three times respectively, are put under vacuum drying chamber, 30 ~ 60 °C and are dried 2 ~ 5 h, are made brick-red
Dendroid Fe2O3；
（2）40 ~ 100 mL, mass fraction are boiled for 0.001 ~ 0.010% chlorauric acid solution, 1 ~ 3 mL, matter is added
The trisodium citrate aqueous solution that fraction is 0.1 ~ 1.0% is measured, 5 ~ 20 min is heated to reflux, treats that solution colour becomes claret,
Solution is cooled to room temperature, is made under solution of gold nanoparticles, 4 °C and is kept in dark place；
（3）By 0.5 ~ 3 mg dendroid Fe2O3It is distributed in 1 mL ultra-pure waters, adds 0.5 ~ 1.5 mL golden nanometer particles
Solution, mixed solution is vibrated at room temperature 12 ~ 32 h, is centrifuged, and removes excessive golden nanometer particle, and Fe is made2O3@
Au, by product Fe2O3@Au are re-dispersed into 1 mL ultra-pure waters, and base material Fe is made2O3@Au solution；
（4）It is 2 ~ 4 mg/mL Fe in 1 ~ 3 mL, concentration2O3In@Au solution, it is 10 μ to add 100 ~ 400 μ L, concentration
G/mL determinand antibody A b1Solution, 24 h of vibration hatching, are centrifuged off excessive determinand antibody A b under 4 °C1, will produce
Thing is distributed in 1 mL, pH 7.4 phosphate buffer solution solution, and antibody A b is made1Capture base material Fe2O3@Au/Ab1
Solution, is stored in standby under 4 °C.
3. it is as claimed in claim 1 a kind of based on dendroid Fe2O3The preparation method of@Au Electrochemiluminescsensor sensor, its
It is characterised by, the secondary antibody Ab2Label Pd-graphene/Fe3O4-Ru(bpy)3 2+/Ab2Solution preparation step is as follows：
（1）It is that 12 mol/L hydrochloric acid solutions are added in 5 ~ 30 mL ultra-pure water by 0.1 ~ 2 mL, concentration, then to solution
0.5 ~ 6.0 g ferric trichlorides solid of middle addition and 0.5 ~ 6.0 g frerrous chloride solids, under continuous stirring, will be mixed molten
Liquid is added dropwise in the sodium hydroxide solution that 100 ~ 500 mL, concentration are 0.5 ~ 5.0 mol/L, 4000 ~
Be centrifuged under 8000 rpm rotating speed, with milli-Q water three times, backward precipitation in add 200 ~ 600 mL, it is dense
Spend for 0.01 ~ 0.04 mol/L hydrochloric acid solution, 5 ~ 15 min of centrifugation under 6000 ~ 9000 rpm rotating speed,
Abandoning supernatant, adds ultra-pure water and carries out the Fe that ultrasound obtains yellow3O4Nano sol；
（2）By 1 ~ 5 mL Fe3O4Nano sol is added to 40 ~ 200 mL, concentration for 1.0 ~ 3.0 mg/mL graphite oxides
In alkene dispersible suspension, under agitation, add 0.10 ~ 0.60 g sodium hydrogensulfite solids, under 80 ~ 160 °C react 2 ~
6 h, are dialysed one week after cooling with ultra-pure water, it is freeze-dried after can obtain the graphene/Fe of 3D structures3O4；
（3）0.06 ~ 0.20 g tetrachloro-palladium acid sodium solids and 0.06 ~ 0.20 g sodium citrate solids are dissolved in 10 ~ 100
In mL ultra-pure waters, it is 0.005 ~ 0.020 mol/L sodium borohydride solutions to add 0.1 ~ 4.0 mL, concentration into solution, is held
30 ~ 90 s of continuous stirring, are made the Pd nano-particle solutions of black；
（4）By 1 ~ 3 mL, the graphene/Fe that concentration is 2 mg/mL3O4The Pd nano-particle solutions of solution and 1 ~ 3 mL
Mixing, lucifuge vibrates 24 ~ 48 h, centrifuges abandoning supernatant, is distributed in 1 mL ultra-pure waters, with 1 ~ 3 mL, concentration
For 1 × 10-3 Mol/L Ru (bpy)3 2+Solution mixing 24 ~ 48 h of vibration, centrifuge abandoning supernatant, are distributed to 1 mL
In ultra-pure water, Pd-graphene/Fe is made3O4-Ru(bpy)3 2+Solution；
（5）It is 2 ~ 4 mg/mL Pd-graphene/Fe in 1 ~ 3 mL, concentration3O4-Ru(bpy)3 2+In solution, 100 are added
~ 400 μ L, concentration are 10 μ g/mL determinand antibody A b2Solution, 24 h of vibration hatching, are centrifuged off excess under 4 °C
Determinand antibody A b2, in the phosphate buffer solution solution that product is distributed to 1 mL, pH 7.4, secondary antibody Ab is made2Mark
Thing Pd-graphene/Fe3O4-Ru(bpy)3 2+/Ab2Solution, is stored in standby under 4 °C.
4. method as claimed in claim 1 prepares a kind of based on dendroid Fe2O3@Au Electrochemiluminescsensor sensor is used to treat
The detection of test sample product, it is characterised in that the Electrochemical Detection that the electrochemical luminescence sensor of the preparation is used for testing sample is walked
It is rapid as follows：
（1）Tested using the three-electrode system of electrochemical workstation, Ag/AgCl electrodes are used as reference electrode, platinum electrode
For to electrode, prepared electrochemical luminescence sensor is working electrode, and electrochemical workstation and chemiluminescence detector are connected
It is connected together, the high pressure of photomultiplier is set to 800 V, cyclic voltammetry scan potential range is 0 ~ 1.2 V, scanning speed
Rate is 0.1 V/s；
（2）In 10 mL, pH 6.4 ~ 8.4 containing the phosphate buffer solution solution that concentration is 35 ~ 65 mmol/L triethylamines
In, by electrochemical luminescence system, the electrochemical luminescence signals intensity produced to the determinand antigen of various concentrations is detected, is drawn
Working curve；
（3）Testing sample solution is detected instead of determinand antigen.
5. one kind as described in one of claim 1-3 is based on dendroid Fe2O3The preparation of@Au Electrochemiluminescsensor sensor
Method, it is characterised in that the determinand antigen is selected from one of following liver cancer marker：AFP, squamous cell carcinoma resist
Former SCCA, CEA, carbohydrate antigen CA15-3.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610803682.9A CN106248657B (en) | 2016-09-05 | 2016-09-05 | One kind is based on dendroid Fe2O3The preparation method and application of@Au Electrochemiluminescsensor sensor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610803682.9A CN106248657B (en) | 2016-09-05 | 2016-09-05 | One kind is based on dendroid Fe2O3The preparation method and application of@Au Electrochemiluminescsensor sensor |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106248657A CN106248657A (en) | 2016-12-21 |
| CN106248657B true CN106248657B (en) | 2017-10-27 |
Family
ID=57599103
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610803682.9A Expired - Fee Related CN106248657B (en) | 2016-09-05 | 2016-09-05 | One kind is based on dendroid Fe2O3The preparation method and application of@Au Electrochemiluminescsensor sensor |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106248657B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109254050B (en) * | 2018-11-05 | 2021-01-12 | 济南大学 | Clenbuterol electrochemiluminescence sensor and preparation method thereof |
| CN109521006B (en) * | 2018-12-24 | 2021-03-19 | 济南大学 | Preparation method and application of double-quenching competitive electrochemiluminescence sensor based on Au @ NiFe MOFs |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101203538A (en) * | 2004-10-15 | 2008-06-18 | 日立化成工业株式会社 | New luminescent compositions and their uses |
| EP2202280A1 (en) * | 2008-11-24 | 2010-06-30 | Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. | Corrosion inhibting coatings controllable by electromagnetic irradiation and methods for corrosion inhibition using the same |
| CN103134793A (en) * | 2013-01-24 | 2013-06-05 | 哈尔滨工业大学 | Electrogenerated chemiluminescence sensor with high sensitivity in cancer cell detection and fabrication method of sensor |
| CN105527427A (en) * | 2016-01-19 | 2016-04-27 | 南昌大学 | Method for fast detecting Listeria monocytogenes |
-
2016
- 2016-09-05 CN CN201610803682.9A patent/CN106248657B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101203538A (en) * | 2004-10-15 | 2008-06-18 | 日立化成工业株式会社 | New luminescent compositions and their uses |
| EP2202280A1 (en) * | 2008-11-24 | 2010-06-30 | Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. | Corrosion inhibting coatings controllable by electromagnetic irradiation and methods for corrosion inhibition using the same |
| CN103134793A (en) * | 2013-01-24 | 2013-06-05 | 哈尔滨工业大学 | Electrogenerated chemiluminescence sensor with high sensitivity in cancer cell detection and fabrication method of sensor |
| CN105527427A (en) * | 2016-01-19 | 2016-04-27 | 南昌大学 | Method for fast detecting Listeria monocytogenes |
Non-Patent Citations (2)
| Title |
|---|
| 含有离子传输单元的聚对苯亚乙烯聚氧化乙烯多嵌段聚合物电致发光材料的合成;朱路等;《应用化学》;20041125(第11期);1151-1155 * |
| 纳米金/碳纳米管复合材料修饰电极催化鲁米诺电致发光体系的研究;黄立漳等;《分析科学学报》;20090220(第01期);47-50 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106248657A (en) | 2016-12-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106442994B (en) | A kind of preparation method and application of the electrochemical immunosensor based on Ag@Au nano composite materials | |
| CN102778561B (en) | Preparation and application of tumor marker immunosensor built by putamen nanometer materials | |
| CN110220888A (en) | A kind of preparation method of the electrochemical luminescence immunosensor of tris (bipyridine) ruthenium functionalization MOF detection Procalcitonin | |
| CN106404756B (en) | One kind is based on graphene/Fe3O4@Au/CeO2/TiO2Electrochemiluminescsensor sensor preparation method and application | |
| Xiang et al. | A redox cycling-amplified electrochemical immunosensor for α-fetoprotein sensitive detection via polydopamine nanolabels | |
| CN109839501B (en) | Electrochemiluminescence immunosensor for measuring circulating tumor cells and preparation method and application thereof | |
| CN102818893A (en) | Preparation and application of Au@Pd core-shell material constructed lung cancer tumor marker immunosensor | |
| CN105572356B (en) | A kind of preparation method and application of breast cancer tumour marker immunosensor | |
| CN105115961B (en) | A kind of preparation method of the electrochemical luminescence sensor of nano composite material | |
| CN107543851B (en) | A kind of preparation method and application of the electrochemical luminescence sensor based on silver oxalate bridging tris (bipyridine) ruthenium nano-complex | |
| CN106596942B (en) | A kind of construction method of interlayer type hepatitis b virus marker immunosensor and application | |
| CN109613244B (en) | Preparation method and application of Ag @ Pt-CuS labeled immunosensor | |
| CN106248657B (en) | One kind is based on dendroid Fe2O3The preparation method and application of@Au Electrochemiluminescsensor sensor | |
| CN108709996A (en) | A kind of preparation method and application of gold-palladium composite Nano enzyme immunosensor | |
| CN104931698A (en) | Preparation method and application of NP-NiGd@Au-based gastric cancer marker gold nano-cluster electrogenerated chemiluminescence sensor | |
| CN111766289A (en) | Oxygen-enriched vacancy CeO2Preparation method and application of electrochemiluminescence immunosensor | |
| CN108896638A (en) | A kind of preparation method and application of the immunosensor based on the graphene-supported sea cucumber shape gold-palladium core-shell nano of titania additive | |
| Li et al. | An electrochemical immunosensor coupling a bamboo-like carbon nanostructure substrate with toluidine blue-functionalized Cu (ii)-MOFs as signal probes for a C-reactive protein assay | |
| CN105842460A (en) | Preparation method of electro-chemiluminescence immunosensor based on silver-hybridized bismuth sulfide | |
| CN104865300B (en) | A kind of based on AuTiO2/Bi2S3The preparation method of the Optical Electro-Chemistry sensor of modified electrode and application | |
| CN112129820B (en) | Construction method and application of specific electrochemical sensor for HER2 detection | |
| Zheng et al. | A Label-Free Immunosensor for CEA based on Pd–Ir bimetallic nanoparticles | |
| CN108375612A (en) | A kind of method of composite nano materials Electrochemical Detection alpha-fetoprotein | |
| CN110702758B (en) | Method for enhancing luminous intensity of squamous cell carcinoma antigen in electrochemical luminescence detection | |
| CN105510599B (en) | A kind of Ru SiO2The preparation method and application for the immunosensor that@PEI nano-particles are built |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into 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 | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20171027 Termination date: 20210905 |