CN106323952B - A kind of preparation method and application of the electroluminescent immunosensor based on double coreagent amplified signals - Google Patents
A kind of preparation method and application of the electroluminescent immunosensor based on double coreagent amplified signals Download PDFInfo
- Publication number
- CN106323952B CN106323952B CN201610935907.6A CN201610935907A CN106323952B CN 106323952 B CN106323952 B CN 106323952B CN 201610935907 A CN201610935907 A CN 201610935907A CN 106323952 B CN106323952 B CN 106323952B
- Authority
- CN
- China
- Prior art keywords
- pct
- mos
- cds
- electrode
- gce
- 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
- G01N27/3278—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction involving nanosized elements, e.g. nanogaps or nanoparticles
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Pathology (AREA)
- Hematology (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Plasma & Fusion (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Nanotechnology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Electrochemistry (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The present invention relates to electrochemiluminescimmunosensor immunosensor technical fields, more particularly to one kind with cadmium sulfide and molybdenum disulfide nano compound (CdS/MoS2) it is luminescent material and base material, it take potassium peroxydisulfate and hydrogen peroxide as the preparation method and application for the immunosensor that double coreagents enhance luminous intensity.By CdS and MoS2Semiconductor nano material similar in two kinds of band gap is compound, and electrical efficiency and electron-hole separative efficiency can be improved；K2S2O8And H2O2Synergistic effect can be played as double coreagents, improves luminous intensity, the enhancing stability of sensor.Based on the specific binding between antigen-antibody, which hinders degree different electron transmission for detecting procalcitonin (PCT), according to the PCT of various concentration, so that sensor electrogenerated chemiluminescence intensity is different, realizes the detection of PCT.
Description
Technical field
The present invention relates to electrochemiluminescimmunosensor immunosensor technical fields, more particularly to one kind with cadmium sulfide and two sulphur
Change molybdenum nano-complex (CdS/MoS2) it is luminescent material and base material, it is double coreagents with potassium peroxydisulfate and hydrogen peroxide
Immunosensor preparation method and application.By CdS and MoS2Semiconductor nano material similar in two kinds of band gap is compound can be with
It effectively improves the luminous intensity of sensor and enhances stability.Potassium peroxydisulfate and hydrogen peroxide do coreagent simultaneously, can be with
Synergistic effect is played, the luminous intensity of sensor is improved.Based on the specific binding between antigen-antibody, the sensor is for examining
It surveys procalcitonin (PCT), hinders degree different electron transmission according to the PCT of various concentration, then make sensor electroluminescentization
It is different to learn luminous intensity, realizes the detection of PCT.
Background technique
PCT belongs to peptide material before a kind of calcitonin, and no hormonal activity, horizontal increase in serum is due to serious thin
Bacterium, fungi, the infection destruction of helminth and pyemia and multiple organ failure and cause, PCT is in autoimmunity, allergy and disease
When poison infection, content will not be increased.Therefore PCT can make diagnosis to bacterium, fungus-caused systemic infection, and PCT will
It is considered as systemic bacterial infections and pyemia auxiliary and the conventional index of antidiastole.At present to the detection hand of PCT antigen
Section mainly have radioimmunology analytic approach, double antibodies sandwich immunochemiluminescence method (ILMA), colloidal gold colorimetric method, transmission be immunized it is turbid
Degree method.The sensitivity with higher of these methods and selectivity, but detection process needs expensive special instrument, needs complexity
Pre-treatment and it is cumbersome, sample consumption is larger, be not suitable for quickly detecting.Electrochemiluminescimmunosensor immunosensor has equipment
Simply, easy to operate, high sensitivity, selectivity is good, detects the advantages that speed is fast.The present invention is based on the immune biographies of electrogenerated chemiluminescence
The advantages of sensor, is prepared for a kind of PCT electrochemiluminescimmunosensor immunosensor based on double coreagent amplified signals, will be immunized
Method is combined with electrogenerated chemiluminescence technology, and the Sensitive Detection of PCT is realized by the specific binding between antigen-antibody.
CdS/MoS in the present invention2As base material, on the one hand since its large specific surface area, film forming ability are strong, Ke Yigu
Determine lot of antibodies；On the other hand due to MoS2There is similar bandgap structure with CdS, the compound photoelectricity that can effectively enhance of the two turns
Efficiency is changed, improves the strength and stability of electrogenerated chemiluminescence, and then improve sensitivity.And potassium peroxydisulfate and hydrogen peroxide do it is double
Coreagent can play synergistic effect, further increase the sensitivity of sensor.The electrogenerated chemiluminescence constructed in the present invention
The advantages that immunosensor has preparation process simple, at low cost, and stability is good, high sensitivity provides one kind for PCT detection
Feasible method.
Summary of the invention
The purpose of the present invention is with CdS/MoS2For substrate and luminescent material, done with potassium peroxydisulfate and hydrogen peroxide double anti-altogether
It answers agent to enhance electroluminescent intensity, constructs a kind of simple and quick, stability is good, the electrogenerated chemiluminescence immune sensing of high sensitivity
Device, and the electrochemiluminescimmunosensor immunosensor is used for the Sensitive Detection of PCT.
The technical solution of the present invention is as follows:
1. a kind of preparation method of the electrochemiluminescimmunosensor immunosensor based on double coreagent amplified signals:
(1) with 1.0 μm, 0.3 μm, 0.05 μm of Al2O3Polishing powder successively polish diameter be 4mm glass-carbon electrode (GCE),
It is cleaned by ultrasonic 5min in ethyl alcohol and ultrapure water respectively, is dried with nitrogen, takes 6~10 μ L CdS/MoS2Nanocomposite dispersion
Drop is coated onto electrode surface, dries film forming at room temperature, is rinsed with PBS (pH 7.4) buffer solution to remove the CdS/ not being bonded
MoS2Obtain CdS/MoS2/GCE；
(2) take procalcitonin antibody (anti-PCT) the standard solution drop coating of 10 μ L, 8~10 μ g/mL to electrode surface, 4
Incubated overnight at DEG C rinses to obtain anti-PCT/CdS/MoS with PBS (pH 7.4) buffer solution2/GCE；
(3) take bovine serum albumin(BSA) (BSA) the solution drop coating that 10 μ L mass fractions are 1~3% to electrode surface, at 37 DEG C
Lower hatching 1h, closes the site of non-specific binding, rinses electrode surface with PBS (pH 7.4) buffer solution and obtains BSA/
anti-PCT/CdS/MoS2/GCE；
(4) the procalcitonin standard solution that a series of various concentrations that 10 μ L concentration are 0.0001~10ng/mL are added dropwise is used
It is identified in antibody specificity, is incubated for 60min at 37 DEG C, rinsed electrode surface with PBS (pH 7.4) buffer solution, be made one
Kind is based on CdS/MoS2Electrochemiluminescimmunosensor immunosensor (PCT/BSA/anti-PCT/CdS/MoS2/GCE)。
Above-mentioned CdS/MoS2The preparation of dispersion liquid:
Weigh the CdCl that a certain amount of molar ratio is 1:12And NaMoO4Be added in 100mL water, ultrasound to being completely dissolved,
2.66g thiocarbamide and 1.0g PVP is added, stirs to obtain milky dispersion liquid, dispersion liquid is transferred in 100mL autoclave, 180 DEG C
React 48h, natural cooling.It is washed three times with water and ethyl alcohol (1:1), is dried overnight, obtains CdS/MoS2Powder.Weigh 10mg
CdS/MoS2Powder ultrasonic is distributed in 10mL ultrapure water to get CdS/MoS2Powder dispersion liquid.
Detection:
(1)PCT/BSA/anti-PCT/CdS/MoS2/ GCE is working electrode, and Ag/AgCl electrode is reference electrode, platinum electricity
Extremely to electrode；It is tested using MPI-B type multi-parameter chemiluminescence analysis test macro, the high pressure of photomultiplier tube is set
It is set to 800V, sweep interval is -1.8~0V, scanning speed 100mV/s；
(2) PBS (pH 7.4) buffering for containing 0.1mol/L potassium peroxydisulfate and 8~12mmol/L hydrogen peroxide in 10mL is molten
In the electrolytic cell of liquid, it is a series of not that 0.0001~10ng/mL is detected by MPI-B type multi-parameter chemiluminescence analysis test macro
With the electrogenerated chemiluminescence intensity of the electrode of concentration PCT standard solution, working curve is drawn；
(3) PCT standard solution is replaced to detect testing sample solution.
The invention has the benefit that
(1) present invention is with CdS/MoS2Two-dimensional semiconductor nano material does substrate, and large specific surface area, film forming ability are strong, can
Fixed lot of antibodies；
(2) present invention is CdS/MoS2Luminescent material drop coating to electrode surface, by protein hinder electron transmission to
The mechanism for changing luminous intensity detects PCT, no enzyme, unmarked, greatly simplifies electrode production process；
(3) present invention CdS and MoS2Semiconductor material similar in two kinds of band gap is compound, improve photoelectric conversion efficiency and
Stability effectively enhances electrogenerated chemiluminescence intensity；
(4) present invention does double coreagents with potassium peroxydisulfate and hydrogen peroxide, can play synergistic effect, effectively enhance
Electrogenerated chemiluminescence intensity；
(5) electrochemiluminescimmunosensor immunosensor prepared by the present invention is used for the detection of PCT, easy to operate, the range of linearity
Width, detection limit is low, and simple, quick, the highly sensitive detection to PCT may be implemented.The range of linearity is 0.0001~10ng/mL, inspection
Rising limit is 0.05pg/mL.
Detailed description of the invention:
Fig. 1 is the electrogenerated chemiluminescence intensity map of various concentration PCT；
Fig. 2 is the Linear Fit Chart of opposite the electrogenerated chemiluminescence intensity and lgc of various concentration PCT.
It wherein, is 0,0.0001 by the concentration that 1 to 10 electrogenerated chemiluminescence intensity map respectively represents PCT in Fig. 1,
0.001,0.005,0.01,0.05,0.1,0.5,1,10ng/mL.
Specific embodiment:
For a better understanding of the present invention, below with specific example come the technical solution that the present invention will be described in detail, but this
Invention is not limited thereto.
A kind of preparation method of the electrochemiluminescimmunosensor immunosensor based on double coreagent amplified signals of embodiment 1:
(1) with 1.0 μm, 0.3 μm, 0.05 μm of Al2O3Polishing powder successively polish diameter be 4mm glass-carbon electrode (GCE),
It is cleaned by ultrasonic 5min in ethyl alcohol and ultrapure water respectively, is dried with nitrogen, takes 6 μ L CdS/MoS2Nanocomposite dispersant liquid drop
It is coated onto electrode surface, dries film forming at room temperature, is rinsed with PBS (pH 7.4) buffer solution to remove the CdS/MoS not being bonded2?
To CdS/MoS2/GCE；
(2) take procalcitonin antibody (anti-PCT) the standard solution drop coating of 10 μ L, 10 μ g/mL to electrode surface, at 4 DEG C
Incubated overnight rinses to obtain anti-PCT/CdS/MoS with PBS (pH 7.4) buffer solution2/GCE；
(3) take bovine serum albumin(BSA) (BSA) the solution drop coating that 10 μ L mass fractions are 1% to electrode surface, at 37 DEG C
Hatch 1h, close the site of non-specific binding, rinses electrode surface with PBS (pH 7.4) buffer solution and obtain BSA/anti-
PCT/CdS/MoS2/GCE；
(4) the procalcitonin standard solution that a series of various concentrations that 10 μ L concentration are 0.0001~10ng/mL are added dropwise is used
It is identified in antibody specificity, is incubated for 60min at 37 DEG C, rinsed electrode surface with PBS (pH 7.4) buffer solution, be made one
Kind is based on CdS/MoS2Electrochemiluminescimmunosensor immunosensor (PCT/BSA/anti-PCT/CdS/MoS2/GCE)。
A kind of electrogenerated chemiluminescence based on double coreagents enhancing semiconductor nanometer composite material light intensity of embodiment 2 is immune
The preparation method of sensor:
(1) with 1.0 μm, 0.3 μm, 0.05 μm of Al2O3Polishing powder successively polish diameter be 4mm glass-carbon electrode (GCE),
It is cleaned by ultrasonic 5min in ethyl alcohol and ultrapure water respectively, is dried with nitrogen, takes 8 μ L CdS/MoS2Nanocomposite dispersant liquid drop
It is coated onto electrode surface, dries film forming at room temperature, is rinsed with PBS (pH 7.4) buffer solution to remove the CdS/MoS not being bonded2?
To CdS/MoS2/GCE；
(2) take procalcitonin antibody (anti-PCT) the standard solution drop coating of 10 μ L, 10 μ g/mL to electrode surface, at 4 DEG C
Incubated overnight rinses to obtain anti-PCT/CdS/MoS with PBS (pH 7.4) buffer solution2/GCE；
(3) take bovine serum albumin(BSA) (BSA) the solution drop coating that 10 μ L mass fractions are 2% to electrode surface, at 37 DEG C
Hatch 1h, close the site of non-specific binding, rinses electrode surface with PBS (pH 7.4) buffer solution and obtain BSA/anti-
PCT/CdS/MoS2/GCE；
(4) the procalcitonin standard solution that a series of various concentrations that 10 μ L concentration are 0.0001~10ng/mL are added dropwise is used
It is identified in antibody specificity, is incubated for 60min at 37 DEG C, rinsed electrode surface with PBS (pH 7.4) buffer solution, be made one
Kind is based on CdS/MoS2Electrochemiluminescimmunosensor immunosensor (PCT/BSA/anti-PCT/CdS/MoS2/GCE)。
A kind of electrogenerated chemiluminescence based on double coreagents enhancing semiconductor nanometer composite material light intensity of embodiment 3 is immune
The preparation method of sensor:
(1) with 1.0 μm, 0.3 μm, 0.05 μm of Al2O3Polishing powder successively polish diameter be 4mm glass-carbon electrode (GCE),
It is cleaned by ultrasonic 5min in ethyl alcohol and ultrapure water respectively, is dried with nitrogen, takes 10 μ L CdS/MoS2Nanocomposite dispersant liquid drop
It is coated onto electrode surface, dries film forming at room temperature, is rinsed with PBS (pH 7.4) buffer solution to remove the CdS/MoS not being bonded2?
To CdS/MoS2/GCE；
(2) take procalcitonin antibody (anti-PCT) the standard solution drop coating of 10 μ L, 10 μ g/mL to electrode surface, at 4 DEG C
Incubated overnight rinses to obtain anti-PCT/CdS/MoS with PBS (pH 7.4) buffer solution2/GCE；
(3) take bovine serum albumin(BSA) (BSA) the solution drop coating that 10 μ L mass fractions are 3% to electrode surface, at 37 DEG C
Hatch 1h, close the site of non-specific binding, rinses electrode surface with PBS (pH 7.4) buffer solution and obtain BSA/anti-
PCT/CdS/MoS2/GCE；
(4) the procalcitonin standard solution that a series of various concentrations that 10 μ L concentration are 0.0001~10ng/mL are added dropwise is used
It is identified in antibody specificity, is incubated for 60min at 37 DEG C, rinsed electrode surface with PBS (pH 7.4) buffer solution, be made one
Kind is based on CdS/MoS2Electrochemiluminescimmunosensor immunosensor (PCT/BSA/anti-PCT/CdS/MoS2/GCE)。
The above-mentioned CdS/MoS of embodiment 42The preparation of dispersion liquid weighs the CdCl that a certain amount of molar ratio is 1:12With
NaMoO4It is added in 100mL water, 2.66g thiocarbamide and 1.0g PVP is added to being completely dissolved in ultrasound, stirs to obtain milky dispersion
Dispersion liquid is transferred in 100mL autoclave by liquid, 180 DEG C of reaction 48h, natural cooling.It is washed three times with water and ethyl alcohol (1:1),
It is dried overnight, obtains CdS/MoS2Powder.Weigh 10mg CdS/MoS2Powder ultrasonic is distributed in 10mL ultrapure water to get CdS/
MoS2Powder dispersion liquid.
The detection of embodiment 5PCT
(1)PCT/BSA/anti-PCT/CdS/MoS2/ GCE is working electrode, and Ag/AgCl electrode is reference electrode, platinum electricity
Extremely to electrode；It is tested using MPI-B type multi-parameter chemiluminescence analysis test macro, the high pressure of photomultiplier tube is set
It is set to 800V, sweep interval is -1.8~0V, scanning speed 100mV/s；
(2) contain PBS (pH 7.4) buffer solution of 0.1mol/L potassium peroxydisulfate and 8mmol/L hydrogen peroxide in 10mL
In electrolytic cell, it is dense that a series of differences of 0.0001~10ng/mL are detected by MPI-B type multi-parameter chemiluminescence analysis test macro
It spends PCT standard solution and does not specifically bind the electrogenerated chemiluminescence intensity of the electrode of PCT, acquired results are shown in Fig. 1, according to gained
Peak value (peak value of the electrode of various concentration PCT standard solution be I, do not specifically bind the electrode of PCT peak value be I0) and
The relationship of PCT concentration draws working curve；
(3)ΔI/I0Fig. 2 (Δ I=I-I is seen with the linear relationship of the logarithm (lgc) of PCT concentration0), as shown in Figure 2, PCT
In 0.1pg/mL~10ng/mL concentration range, Δ I/I0Logarithm with PCT concentration is in good linear correlation, linear equation
For Δ I/I0=0.107lgc+0.461, c are concentration, and unit is g/mL, and linearly dependent coefficient 0.981, detection is limited to
0.05pg/mL；
(4) PCT standard solution is replaced to detect testing sample solution.
The detection of embodiment 6PCT
(1)PCT/BSA/anti-PCT/CdS/MoS2/ GCE is working electrode, and Ag/AgCl electrode is reference electrode, platinum electricity
Extremely to electrode；It is tested using MPI-B type multi-parameter chemiluminescence analysis test macro, the high pressure of photomultiplier tube is set
It is set to 800V, sweep interval is -1.8~0V, scanning speed 100mV/s；
(2) contain PBS (pH 7.4) buffer solution of 0.1mol/L potassium peroxydisulfate and 10mmol/L hydrogen peroxide in 10mL
Electrolytic cell in, pass through MPI-B type multi-parameter chemiluminescence analysis test macro detect a series of differences of 0.0001~10ng/mL
Concentration PCT standard solution and do not specifically bind PCT electrode electrogenerated chemiluminescence intensity, acquired results are shown in Fig. 1, according to institute
(peak value of the electrode of various concentration PCT standard solution is denoted as I to the peak value obtained, and the peak value for not specifically binding the electrode of PCT is
I0) and PCT concentration relationship, draw working curve；
(3)ΔI/I0Fig. 2 (Δ I=I-I is seen with the linear relationship of the logarithm (lgc) of PCT concentration0), as shown in Figure 2, PCT
In 0.1pg/mL~10ng/mL concentration range, Δ I/I0Logarithm with PCT concentration is in good linear correlation, linear equation
For Δ I/I0=0.107lgc+0.461, c are concentration, and unit is g/mL, and linearly dependent coefficient 0.981, detection is limited to
0.05pg/mL；
(4) PCT standard solution is replaced to detect testing sample solution.
The detection of embodiment 7PCT
(1)PCT/BSA/anti-PCT/CdS/MoS2/ GCE is working electrode, and Ag/AgCl electrode is reference electrode, platinum electricity
Extremely to electrode；It is tested using MPI-B type multi-parameter chemiluminescence analysis test macro, the high pressure of photomultiplier tube is set
It is set to 800V, sweep interval is -1.8~0V, scanning speed 100mV/s；
(2) contain PBS (pH 7.4) buffer solution of 0.1mol/L potassium peroxydisulfate and 12mmol/L hydrogen peroxide in 10mL
Electrolytic cell in, pass through MPI-B type multi-parameter chemiluminescence analysis test macro detect a series of differences of 0.0001~10ng/mL
Concentration PCT standard solution and do not specifically bind PCT electrode electrogenerated chemiluminescence intensity, acquired results are shown in Fig. 1, according to institute
(peak value of the electrode of various concentration PCT standard solution is denoted as I to the peak value obtained, and the peak value for not specifically binding the electrode of PCT is denoted as
I0) and PCT concentration relationship, draw working curve；
(3)ΔI/I0Fig. 2 (Δ I=I-I is seen with the linear relationship of the logarithm (lgc) of PCT concentration0), as shown in Figure 2, PCT
In 0.1pg/mL~10ng/mL concentration range, Δ I/I0Logarithm with PCT concentration is in good linear correlation, linear equation
For Δ I/I0=0.107lgc+0.461, c are concentration, and unit is g/mL, and linearly dependent coefficient 0.981, detection is limited to
0.05pg/mL；
(4) PCT standard solution is replaced to detect testing sample solution.
Claims (3)
1. a kind of preparation method of the electrochemiluminescimmunosensor immunosensor based on double coreagent amplified signals, feature exist
In, comprising the following steps:
(1) with 1.0 μm, 0.3 μm, 0.05 μm of Al2O3Polishing powder polishes diameter successively as the glass-carbon electrode GCE of 4mm, exists respectively
It is cleaned by ultrasonic 5min in ethyl alcohol and ultrapure water, is dried with nitrogen, takes 6~10 μ L CdS/MoS2Nanocomposite dispersion liquid drop coating
To electrode surface, film forming is dried at room temperature, is rinsed with 7.4 buffer solution of PBS pH to remove the CdS/MoS not being bonded2It obtains
CdS/MoS2/GCE；
(2) it takes the procalcitonin antibody anti-PCT standard solution drop coating of 10 μ L, 8~10 μ g/mL to electrode surface, is incubated at 4 DEG C
Change overnight, rinses to obtain anti-PCT/CdS/MoS with 7.4 buffer solution of PBS pH2/GCE；
(3) it takes the bovine serum albumin(BSA) BSA solution drop coating that 10 μ L mass fractions are 1~3% to electrode surface, hatches at 37 DEG C
1h closes the site of non-specific binding, rinses electrode surface with 7.4 buffer solution of PBS pH and obtains BSA/anti-PCT/
CdS/MoS2/GCE；
(4) be added dropwise 10 μ L concentration be 0.0001~10ng/mL a series of various concentrations procalcitonin standard solution be used for
Antibody specificity identification, is incubated for 60min at 37 DEG C, rinses electrode surface with 7.4 buffer solution of PBS pH, a kind of base is made
In CdS/MoS2Electrochemiluminescimmunosensor immunosensor PCT/BSA/anti-PCT/CdS/MoS2/GCE；
The CdS/MoS2Nanocomposite dispersion liquid, which is characterized in that making step is as follows:
Weigh the CdCl that a certain amount of molar ratio is 1:12And NaMoO4It is added in 100mL water, ultrasound is added to being completely dissolved
2.66g thiocarbamide and 1.0g PVP stir to obtain milky dispersion liquid, dispersion liquid are transferred in 100mL autoclave, 180 DEG C of reactions
48h, natural cooling three times with the water and ethanol washing of 1:1 are dried overnight, obtain CdS/MoS2Powder；Weigh 10mg CdS/MoS2
Powder ultrasonic is distributed in 10mL ultrapure water to get CdS/MoS2Nanocomposite dispersion liquid.
2. application of the electrochemiluminescimmunosensor immunosensor of claim 1 the method preparation in detection procalcitonin.
3. application according to claim 2, which is characterized in that the detecting step for PCT detection is as follows:
(1)PCT/BSA/anti-PCT/CdS/MoS2/ GCE is working electrode, and Ag/AgCl electrode is reference electrode, and platinum electrode is
To electrode；It is tested using MPI-B type multi-parameter chemiluminescence analysis test macro, is set the high pressure of photomultiplier tube to
800V, sweep interval are -1.8~0V, scanning speed 100mV/s；
(2) contain 7.4 buffer solution of PBS pH of 0.1mol/L potassium peroxydisulfate and 8~12mmol/L hydrogen peroxide in 10mL
In electrolytic cell, it is dense that a series of differences of 0.0001~10ng/mL are detected by MPI-B type multi-parameter chemiluminescence analysis test macro
The electrogenerated chemiluminescence intensity of the electrode of PCT standard solution is spent, working curve is drawn；
(3) PCT standard solution is replaced to detect testing sample solution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610935907.6A CN106323952B (en) | 2016-11-01 | 2016-11-01 | A kind of preparation method and application of the electroluminescent immunosensor based on double coreagent amplified signals |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610935907.6A CN106323952B (en) | 2016-11-01 | 2016-11-01 | A kind of preparation method and application of the electroluminescent immunosensor based on double coreagent amplified signals |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106323952A CN106323952A (en) | 2017-01-11 |
| CN106323952B true CN106323952B (en) | 2019-04-02 |
Family
ID=57818806
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610935907.6A Expired - Fee Related CN106323952B (en) | 2016-11-01 | 2016-11-01 | A kind of preparation method and application of the electroluminescent immunosensor based on double coreagent amplified signals |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106323952B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110231335A (en) * | 2019-06-18 | 2019-09-13 | 济南大学 | A kind of preparation method and application based on ferritin three-dimensional structure immunosensor |
| CN110441297B (en) * | 2019-08-29 | 2021-08-20 | 济南大学 | Preparation method of electrochemical luminescence sensor based on tetraphenyl ethylene nanocluster |
| CN110794017B (en) * | 2019-10-29 | 2022-06-10 | 济南大学 | Preparation method of electrochemical immunosensor for detecting procalcitonin |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105759026A (en) * | 2016-03-07 | 2016-07-13 | 济南大学 | Preparation method and application of simple electrochemiluminescence immunosensor based on compound semiconductor nanomaterial |
-
2016
- 2016-11-01 CN CN201610935907.6A patent/CN106323952B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN106323952A (en) | 2017-01-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Chen et al. | A gold nanoparticles/sol–gel composite architecture for encapsulation of immunoconjugate for reagentless electrochemical immunoassay | |
| CN106324060A (en) | Preparation method and application of PCT electrochemical immunosensor based on AuNPs/Cu-MOF marking | |
| CN103116023B (en) | ECL (electrochemiluminescence) immunosensor for detecting tumor markers and preparation method and applications thereof | |
| Wu et al. | Bioconjugated persistent luminescence nanoparticles for Föster resonance energy transfer immunoassay of prostate specific antigen in serum and cell extracts without in situ excitation | |
| Patris et al. | Antibodies as target for affinity biosensors | |
| CN106568973A (en) | Preparation method and application of MoS2/Au-Pd compound-based procalcitonin electrochemical immunosensor | |
| CN106323952B (en) | A kind of preparation method and application of the electroluminescent immunosensor based on double coreagent amplified signals | |
| CN102507953A (en) | Preparation method of electrochemistry immunosensor for determining alpha fetoprotein | |
| CN105044083A (en) | Method for preparing electrochemiluminescence immunosensor for alpha fetoprotein based on Au-g-C3N4 nanocomposite and application | |
| CN101655473B (en) | Preparation method of nanogold immunoelectrode | |
| CN107389949A (en) | A kind of electrochemical immunosensor preparation method for PCSK9 Protein Detections | |
| CN101943703A (en) | Nanotechnology-based trace protein detection method | |
| CN104391123B (en) | A kind of preparation method of the biology sensor built based on flower-like nanometer ZnO microsphere and golden palladium nano flower composite material and application | |
| Joung et al. | Ultra-sensitive detection of pathogenic microorganism using surface-engineered impedimetric immunosensor | |
| CN104459124A (en) | Preparation method and application of electrochemical immunosensor based on HS-beta-CD-Ag-GOD conjugate | |
| Nie et al. | A highly sensitive capillary-based immunosensor by combining with peroxidase nanocomplex-mediated signal amplification for detection of procalcitonin in human serum | |
| CN106066324A (en) | A kind of preparation method and application of electroluminescent chemiluminescence biosensor label | |
| CN111707721B (en) | Preparation method and application of 17 beta-estradiol electrochemiluminescence aptamer sensor | |
| CN110441297A (en) | A kind of electrochemical luminescence quenching type transducer production method and application based on tetraphenyl ethylene nano-cluster | |
| CN109115855A (en) | A kind of preparation method and application for the electrochemical immunosensor detecting Alzheimer's disease marker | |
| CN105806908A (en) | Preparation method and application of label-free type electrochemical immunosensor based on MoS2/Au composite material | |
| CN105759026A (en) | Preparation method and application of simple electrochemiluminescence immunosensor based on compound semiconductor nanomaterial | |
| Zhong et al. | Multiplex immunoassay of chicken cytokines via highly-sensitive chemiluminescent imaging array | |
| CN105842460A (en) | Preparation method of electro-chemiluminescence immunosensor based on silver-hybridized bismuth sulfide | |
| CN105606681A (en) | Preparation method and application of biosensor built based on gold and copper-multiwalled carbon nanotube-manganese dioxide |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | 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: 20190402 Termination date: 20201101 |