CN109507174A - Preparation based on the compound ZnO nanoparticle quenching Particles in Electrochemiluminescofce ofce Luminol sensor of curcumin - Google Patents
Preparation based on the compound ZnO nanoparticle quenching Particles in Electrochemiluminescofce ofce Luminol sensor of curcumin Download PDFInfo
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Abstract
The present invention relates to a kind of golden hydridization MoS2/Bi2S3The electrochemical luminescence sensor of the immobilized luminol detection amyloid beta protein 42 of nanometer rods.In the present invention, MoS2/Bi2S3Nanometer rods not only have excellent electrocatalysis characteristic, but also can enhance the electrochemical luminescence intensity of luminescent material by the immobilized a large amount of luminescent material luminol of gold-sulfide linkage.In order to delicately detect amyloid beta protein 42, the present invention devises a kind of quenching type electrochemical luminescence immunosensor of interlayer type, using curcumin-ZnO nano material based on consumption super oxygen root free radical and electrochemical luminescence-Resonance energy transfer quenching luminol electrochemical luminescence signals.It can be in conjunction with different amounts of secondary antibody marker poly-dopamine@curcumin-ZnO, so that the sensor electrochemical luminescence Strength Changes are different according to the amyloid beta protein 42 of various concentration.The present invention is 0.05 pg/mL-10 ng/mL to the range of linearity that amyloid beta protein 42 detects, and detection is limited to 21 fg/mL.
Description
Technical field
The present invention relates to a kind of golden hydridization MoS2/Bi2S3The electrification of the immobilized luminol detection amyloid beta protein 42 of nanometer rods
Learn luminescence sensor.Specifically using golden hydridization MoS2/Bi2S3For the immobilized luminol of nanometer rods as luminescent material, curcumin is compound
ZnO nanoparticle prepares a kind of 42 quenching type electrochemical luminescence sensor of detection amyloid beta protein, belongs to electricity as quencher
Chemiluminescence detection technology field.
Background technique
Alzheimer disease is a kind of nervous system degenerative disease for betiding old age and presenium, and clinical manifestation is
Memory disorders, execute dysfunction and personality and behavior change etc. at aphasia.Therefore, Alzheimer disease seriously threatens people
The health of class, and reduce the quality of life of the mankind.The pathogenesis of the deposition Ahl tribulus sea silent sickness of amyloid beta protein has
Certain relationship.Amyloid beta protein 42 and amyloid beta protein 40 are two kinds of main components of amyloid beta protein, wherein starch
Main component in sample β protein 42 Alzheimer disease patient's patch is easier to assemble than amyloid beta protein 40.Early detection,
Early treatment will improve the life quality of patient.It therefore, in the present invention, is test object with amyloid beta protein 42, exploitation
A kind of novel, sensitive method of immunity, is significantly.
Electrochemical luminescence (ECL) analysis has high sensitivity, and the range of linearity is wide；It is good to react controllability, space-time controllability；Instrument
Device is simple, and analysis speed is fast；Save reagent；Analysis has a wide range of application；Much information can be obtained simultaneously, and it is fast to be conducive to research
The advantages such as fast luminescence-producing reaction and luminescence-producing reaction mechanism have been developed as a subdiscipline of analytical chemistry.Luminol conduct
Traditional electrochemical luminescence reagent has efficient luminous efficiency, however, how luminol is steadily immobilized on electrode surface
Solid-state ECL sensor is constructed, it is most important for the stability and sensitivity that solve ECL sensor.It is well known that unmarked type
Sensor causes the variation of ECL signal to realize the detection of object the hindrance function of modified electrode according to biomolecule.Therefore,
It was found that novel quencher causes the variation of luminescent material ECL signal by energy transfer, for realizing that the trace of object is examined
Survey has great importance.
Summary of the invention
The present invention devises a kind of electrochemical luminescence immunosensor of quenching type for detecting amyloid beta protein.
In the present invention, using golden hydridization MoS2/Bi2S3The immobilized luminescent material luminol of nanometer rods, significantly enhances Shandong
The ECL intensity of minot.MoS2/Bi2S3Nanometer rods not only have certain catalytic action to the decomposition of hydrogen peroxide, but also can be with
More gold nanoparticles are closed by Au-S bond, to pass through gold-NH2The immobilized more luminols of key and amyloid beta protein
42 antibody.The present invention uses MoS2/Bi2S3Nanometer rods utilize MoS as base material2And Bi2S3Between the hetero-junctions that is formed change
Its electrochemical properties has been apt to it, with simple MoS2And Bi2S3Nano material is compared, and the immobilized luminol of composite nano materials has steady
Fixed and strong electrochemical luminescence signals, improve the stability and sensitivity of sensor.In order to delicately detect amyloid beta protein
42, the compound ZnO nanoparticle of curcumin reduces the electrochemical luminescence intensity of luminol as quencher.Antioxidant curcumin
It can be with super oxygen root free radical (O2 •−) reaction, and the electrochemical luminescence intensity and O of luminol2 •−Content it is directly proportional, because of O2 •−
It is to generate one of luminol free radical and the essential substance of excitation state 3- aminophthalic acid salt.Also, curcumin
Ultraviolet-ray visible absorbing peak has certain wave spectrum Chong Die with the fluorescence emission peak of luminol, i.e., there is also certain therebetween
Energy transfer has further quenched the electrochemical luminescence intensity of luminol.The hydrophobicity of curcumin limits it in bioanalysis
Curcumin is fixed in nano material the dispersibility for improving it in water by the application in field, widens it in biosensor
The application in field.In the present invention, curcumin is compound with ZnO nanoparticle, secondary antibody, poly- DOPA are simply combined in order to stablize
Amine auto polymerization in composite material surface, by Michael react the benzoquinones functional group in poly-dopamine can in biomolecule
Amino formed covalent bond.The amyloid beta protein 42 of various concentration can combine different amounts of two anti-poly-dopamine curcumins-
ZnO, so as to cause luminol-gold hydridization MoS2/Bi2S3Amyloid beta protein 42 is realized in the variation of nanometer rods luminous intensity
Detection.
To achieve the goals above, The technical solution adopted by the invention is as follows:
1. a kind of gold hydridization MoS2/Bi2S3The electrochemical luminescence sensing of the immobilized luminol detection amyloid beta protein 42 of nanometer rods
Device, preparation step are as follows:
(1) using the glass-carbon electrode of polishing powder pretreatment 4 mm of diameter, ultrapure water is clean；
(2) by 7 μ L, 10 mg/mL luminol-gold hydridization MoS2/Bi2S3Nanometer rods solution drop coating is to electrode surface, room temperature preservation
To drying；
(3) 6 μ L of drop coating, 500 μ g/mL primary antibody Ab1Solution is saved in glassy carbon electrode surface, 4 DEG C of refrigerators to drying, ultrapure
Water cleaning；
(4) the bovine serum albumin(BSA) BSA that 3 μ L mass fraction of drop coating is 1% closes nonspecific activity site, in 4 DEG C of refrigerators
It saves to drying, ultrapure water cleaning；
(5) 42 drop coating of amyloid beta protein of 6 μ L various concentrations is saved in electrode surface, 4 DEG C of refrigerators to drying, it is ultrapure
Water cleaning；
(6) by 6 μ L, 1~7 mg/mL Ab2Poly-dopamine@curcumin-ZnO solution drop coating is in electrode surface, 4 DEG C of refrigerators
It saves to drying, ultrapure water cleaning obtains the electrochemical luminescence biosensor of detection amyloid beta protein 42.
2. a kind of golden hydridization MoS of the present invention2/Bi2S3The immobilized luminol detection amyloid beta protein 42 of nanometer rods
Electrochemical luminescence sensor, the luminol-gold hydridization MoS2/Bi2S3Nanometer rods solution, preparation step are as follows:
(1) MoS2/Bi2S3The preparation of nanometer rods
By 0.242 g Na2MoO4·2H2O and 0.765 g Bi (NO3)3·5H2O is dispersed in 60 mL ultrapure waters, is added 0.76
G thiocarbamide, 1 h of magnetic agitation.Above-mentioned solution is transferred in 100 mL reaction kettles, 220 DEG C of 24 h of reaction, centrifugation, is done washing
It is dry to obtain MoS2/Bi2S3Nanometer rods；
(2) luminol-gold hydridization MoS2/Bi2S3The preparation of nanometer rods
By the MoS of preparation2/Bi2S3Nanometer rods are dispersed in 50 mL ultrapure waters, 1 h of ultrasound.Then, by 2 mL, 1% HAuCl4
It is added in above-mentioned solution with 5 mg PVP, after stirring 6 h, 2 mL, 50 mmol/L sodium citrate solution is added dropwise and lacks
The NaBH of amount4Solution reduction HAuCl4.After stirring 6 h, the unbonded gold nanoparticle of centrifugation removal obtains golden hydridization
MoS2/Bi2S3Nanometer rods.Then nanometer rods are dispersed in 5 mL ultrapure waters, 1~5 mL 5mmol/L luminol is added and stays overnight
Stirring, passes through gold-NH2Luminol is incorporated in nano-bar material surface by key, and the unbonded luminol of centrifuge separation removal obtains
Luminol-gold hydridization MoS2/Bi2S3Nanometer rods.
3. a kind of golden hydridization MoS of the present invention2/Bi2S3The immobilized luminol detection amyloid beta protein 42 of nanometer rods
Electrochemical luminescence sensor, the Ab2Poly-dopamine@curcumin-ZnO, preparation step are as follows:
(1) preparation of curcumin-ZnO
5 mg curcumins are dispersed in 50 mL ultrapure waters, 90 DEG C are back to curcumin and are completely dissolved.50 mL 0.1 are added
Mol/L zinc nitrate solution, 90 DEG C of 1 h of reflux.When above-mentioned solution is cooled to room temperature, 5 mL, 0.2 mol/L is added in ice-water bath
KOH stirs 1 h, forms orange-yellow colloidal suspensions, with the unbonded curcumin of ultrapure water and acetone washing removal, vacuum
It is dried to obtain curcumin-ZnO；
(2) Ab2Poly-dopamine@curcumin-ZnO
Curcumin-the ZnO of 20 mg preparation and 1~5 mg dopamine are dispersed in 30 mL ultrapure waters, 6 h are stirred, centrifugation is gone
Except unbonded dopamine.Said mixture is dispersed in 10 Tris-HCl(pH=8.5 mmol/L 10 mL) in solution, stir
Mix 6 h, the unbonded poly-dopamine of centrifugation removal.Then, 1~10 mg poly-dopamine@curcumin-ZnO is dispersed in 5 mL phosphorus
In hydrochlorate buffer solution (pH=7.4), 500 μ L, 500 μ g/mL secondary antibody Ab is added2, 6 h are stirred under 4 C.Later, 100 are added
1% bovine serum albumin(BSA) of μ L, closes non-specific sites, and centrifugation obtains Ab2Poly-dopamine@curcumin-ZnO, is dispersed 1
ML is stored in 4 DEG C of refrigerators stand-by in phosphate buffer solution (pH=7.4).
4. preparation method of the present invention preparation based on luminol-gold hydridization MoS2/Bi2S3Nanometer rods building electricity
Chemiluminescence sensor, for the detection of amyloid beta protein 42, steps are as follows:
(1) using reference electrode-Ag/AgCl electrode, to electrode-platinum electrode, obtained electrochemical luminescence sensor as work
Make electrode, be connected in the magazine of chemiluminescence detector, electrochemical workstation and chemiluminescence detector are linked together,
The high pressure of photomultiplier tube is set as 600 V, and scanning voltage is set as -0.2~0.6 V；
(2) it using the PBS buffer solution of the mmol/L hydrogen peroxide Han 1 mmol/L~8, is detected by Electrochemiluminescince different
The electrochemical luminescence signals intensity that the amyloid beta protein 42 of concentration generates；
The PBS buffer solution, pH=6.5~8.5 are with 1/15 mol/L Na2HPO4With 1/15 mol/L KH2PO4Match
System；
(3) according to the linear relationship of resulting electrochemical luminescence intensity value and 42 log concentration of amyloid beta protein, it is bent to draw work
Line.
Detailed description of the invention
Fig. 1 is the process design drawing for the sensor that the present invention designs.
Specific embodiment
Present invention will be further explained below with reference to specific examples.It should be understood that these embodiments are merely to illustrate the present invention
Rather than it limits the scope of the invention.
Embodiment 1 prepares luminol-gold hydridization MoS2/Bi2S3Nanometer rods solution
(1) MoS2/Bi2S3The preparation of nanometer rods
By 0.242 g Na2MoO4·2H2O and 0.765 g Bi (NO3)3·5H2O is dispersed in 60 mL ultrapure waters, is added 0.76
G thiocarbamide, 1 h of magnetic agitation.Above-mentioned solution is transferred in 100 mL reaction kettles, 220 DEG C of 24 h of reaction, centrifugation, is done washing
It is dry to obtain MoS2/Bi2S3Nanometer rods；
(2) luminol-gold hydridization MoS2/Bi2S3The preparation of nanometer rods
By the MoS of preparation2/Bi2S3Nanometer rods are dispersed in 50 mL ultrapure waters, 1 h of ultrasound.By 2 mL, 1% HAuCl4With 5 mg
PVP is added in above-mentioned solution, and after stirring 6 h, 2 mL, 50 mmol/L sodium citrate solution and a small amount of is added dropwise
NaBH4Solution reduction HAuCl4.After stirring 6 h, the unbonded gold nanoparticle of centrifugation removal obtains the MoS of golden hydridization2/
Bi2S3Nanometer rods.Then nanometer rods are dispersed in 5 mL ultrapure waters, 5 mL 5mmol/L luminols is added and are stirred overnight, pass through
Gold-NH2Luminol is incorporated in nano-bar material surface by key, and the unbonded luminol of centrifuge separation removal obtains luminol-gold
The MoS of hydridization2/Bi2S3Nanometer rods.
Embodiment 2 prepares Ab2Poly-dopamine@curcumin-ZnO
(1) preparation of curcumin-ZnO
5 mg curcumins are dispersed in 50 mL ultrapure waters, 90 DEG C are back to curcumin and are completely dissolved.50 mL 0.1 are added
Mol/L zinc nitrate solution, 90 DEG C of 1 h of reflux.When above-mentioned solution is cooled to room temperature, 5 mL, 0.2 mol/L is added in ice-water bath
KOH stirs 1 h, forms orange-yellow colloidal suspensions, with the unbonded curcumin of ultrapure water and acetone washing removal, vacuum
It is dried to obtain curcumin-ZnO；
(2) Ab2Poly-dopamine@curcumin-ZnO
Curcumin-the ZnO of 20 mg preparation and 1 mg dopamine are dispersed in 30 mL ultrapure waters, stir 6 h, centrifugation removal is not
In conjunction with dopamine.Said mixture is dispersed in 10 Tris-HCl(pH=8.5 mmol/L 10 mL) in solution, stirring 6
H, the unbonded poly-dopamine of centrifugation removal.Then, 1 mg poly-dopamine@curcumin-ZnO is dispersed in 5 mL phosphate-buffereds
In solution (pH=7.4), 500 μ L, 500 μ g/mL secondary antibody Ab is added2, 6 h are stirred under 4 C.Later, 1% N of 100 μ L is added
Seralbumin, closes non-specific sites, and centrifugation obtains Ab2Poly-dopamine@curcumin-ZnO, is dispersed in 1 mL phosphorus
In hydrochlorate buffer solution (pH=7.4), it is stored in 4 DEG C of refrigerators stand-by.
Embodiment 3 prepares Ab2Poly-dopamine@curcumin-ZnO
(1) preparation of curcumin-ZnO
5 mg curcumins are dispersed in 50 mL ultrapure waters, 90 DEG C are back to curcumin and are completely dissolved.Then, 50 mL are added
0.1 mol/L zinc nitrate solution, 90 DEG C of 1 h of reflux.When above-mentioned solution is cooled to room temperature, 5 mL 0.2 are added in ice-water bath
Mol/L KOH stirs 1 h, forms orange-yellow colloidal suspensions, with the unbonded turmeric of ultrapure water and acetone washing removal
Element, vacuum drying obtain curcumin-ZnO；
(2) Ab2Poly-dopamine@curcumin-ZnO
Curcumin-the ZnO of 20 mg preparation and 5 mg dopamines are dispersed in 30 mL ultrapure waters, stir 6 h, centrifugation removal is not
In conjunction with dopamine.Said mixture is dispersed in 10 Tris-HCl(pH=8.5 mmol/L 10 mL) in solution, stirring 6
H, the unbonded poly-dopamine of centrifugation removal.Then, 5 mg poly-dopamine@curcumin-ZnO are dispersed in 5 mL phosphate-buffereds
In solution (pH=7.4), 500 μ L, 500 μ g/mL secondary antibody Ab is added2, 6 h are stirred under 4 C.Later, 1% N of 100 μ L is added
Seralbumin, closes non-specific sites, and centrifugation obtains Ab2Poly-dopamine@curcumin-ZnO, is dispersed in 1 mL phosphorus
In hydrochlorate buffer solution (pH=7.4), it is stored in 4 DEG C of refrigerators stand-by.
Embodiment 4 prepares Ab2Poly-dopamine@curcumin-ZnO
(1) preparation of curcumin-ZnO
5 mg curcumins are dispersed in 50 mL ultrapure waters, 90 DEG C are back to curcumin and are completely dissolved.Then, 50 mL are added
0.1 mol/L zinc nitrate solution, 90 DEG C of 1 h of reflux.When above-mentioned solution is cooled to room temperature, 5 mL 0.2 are added in ice-water bath
Mol/L KOH stirs 1 h, forms orange-yellow colloidal suspensions, with the unbonded turmeric of ultrapure water and acetone washing removal
Element, vacuum drying obtain curcumin-ZnO；
(2) Ab2Poly-dopamine@curcumin-ZnO
Curcumin-the ZnO of 20 mg preparation and 10 mg dopamines are dispersed in 30 mL ultrapure waters, 6 h, centrifugation removal are stirred
Unbonded dopamine.Said mixture is dispersed in 10 Tris-HCl(pH=8.5 mmol/L 10 mL) in solution, stirring 6
H, the unbonded poly-dopamine of centrifugation removal.Then, 10 mg poly-dopamine@curcumin-ZnO 5 mL phosphate are dispersed in delay
It rushes in solution (pH=7.4), 500 μ L, 500 μ g/mL secondary antibody Ab is added2, 6 h are stirred under 4 C.Later, 100 μ L 1% are added
Bovine serum albumin(BSA), closes non-specific sites, and centrifugation obtains Ab2Poly-dopamine@curcumin-ZnO, is dispersed in 1 mL
In phosphate buffer solution (pH=7.4), it is stored in 4 DEG C of refrigerators stand-by.
The electrochemical luminescence sensor of the preparation detection amyloid beta protein 42 of embodiment 5
(1) using the glass-carbon electrode of polishing powder pretreatment 4 mm of diameter, ultrapure water is clean；
(2) by 7 μ L, 10 mg/mL luminol-gold hydridization MoS2/Bi2S3Nanometer rods solution drop coating is to electrode surface, room temperature preservation
To drying；
(3) 6 μ L of drop coating, 500 μ g/mL primary antibody Ab1Solution is saved in glassy carbon electrode surface, 4 DEG C of refrigerators to drying, ultrapure
Water cleaning；
(4) bovine serum albumin(BSA) that 3 μ L mass fraction of drop coating is 1% is closed nonspecific activity site, is saved in 4 DEG C of refrigerators
To drying, ultrapure water cleaning；
(5) 42 drop coating of amyloid beta protein of 6 μ L various concentrations is saved in electrode surface, 4 DEG C of refrigerators to drying, it is ultrapure
Water cleaning；
(6) by 6 μ L, 3 mg/mL Ab2Poly-dopamine@curcumin-ZnO solution drop coating is protected in electrode surface, 4 DEG C of refrigerators
It deposits to drying, ultrapure water cleaning obtains the electrochemical luminescence biosensor of detection amyloid beta protein 42.
The electrochemical luminescence sensor of the preparation detection amyloid beta protein 42 of embodiment 6
(1) using the glass-carbon electrode of polishing powder pretreatment 4 mm of diameter, ultrapure water is clean；
(2) by 7 μ L, 10 mg/mL luminol-gold hydridization MoS2/Bi2S3Nanometer rods solution drop coating is to electrode surface, room temperature preservation
To drying；
(3) 6 μ L of drop coating, 500 μ g/mL primary antibody Ab1Solution is saved in glassy carbon electrode surface, 4 DEG C of refrigerators to drying, ultrapure
Water cleaning；
(4) bovine serum albumin(BSA) that 3 μ L mass fraction of drop coating is 1% is closed nonspecific activity site, is saved in 4 DEG C of refrigerators
To drying, ultrapure water cleaning；
(5) 42 drop coating of amyloid beta protein of 6 μ L various concentrations is saved in electrode surface, 4 DEG C of refrigerators to drying, it is ultrapure
Water cleaning；
(6) by 6 μ L, 7 mg/mL Ab2Poly-dopamine@curcumin-ZnO solution drop coating is protected in electrode surface, 4 DEG C of refrigerators
It deposits to drying, ultrapure water cleaning obtains the electrochemical luminescence biosensor of detection amyloid beta protein 42.
The detection method of 7 amyloid beta protein 42 of embodiment
(1) using reference electrode-Ag/AgCl electrode, to electrode-platinum electrode, obtained electrochemical luminescence sensor as work
Make electrode, be connected in the magazine of chemiluminescence detector, electrochemical workstation and chemiluminescence detector are linked together,
The high pressure of photomultiplier tube is set as 600 V, and scanning voltage is set as -0.2~0.6 V；
(2) using the PBS buffer solution for containing 3 mmol/L hydrogen peroxide, the starch of various concentration is detected by Electrochemiluminescince
The electrochemical luminescence signals intensity that sample β protein 42 generates；
The PBS buffer solution, pH=6.5 are with 1/15 mol/L Na2HPO4With 1/15 mol/L KH2PO4It prepares；
(3) according to the linear relationship of resulting electrochemical luminescence intensity value and 42 log concentration of amyloid beta protein, it is bent to draw work
Line.
The detection method of 8 amyloid beta protein 42 of embodiment
(1) using reference electrode-Ag/AgCl electrode, to electrode-platinum electrode, obtained electrochemical luminescence sensor as work
Make electrode, be connected in the magazine of chemiluminescence detector, electrochemical workstation and chemiluminescence detector are linked together,
The high pressure of photomultiplier tube is set as 600 V, and scanning voltage is set as -0.2~0.6 V；
(2) using the PBS buffer solution for containing 5 mmol/L hydrogen peroxide, the starch of various concentration is detected by Electrochemiluminescince
The electrochemical luminescence signals intensity that sample β protein 42 generates；
The PBS buffer solution, pH=8.0 are with 1/15 mol/L Na2HPO4With 1/15 mol/L KH2PO4It prepares；
(3) according to the linear relationship of resulting electrochemical luminescence intensity value and 42 log concentration of amyloid beta protein, it is bent to draw work
Line.
The detection of 9 synthetic cerebrospinal fluid amyloid beta protein 42 of embodiment
(1) amyloid beta protein 42 of various concentration is added into diluted synthetic cerebrospinal fluid, using Standard Addition Method for Determination sample
The average recovery rate of middle amyloid beta protein 42, the results are shown in Table 1.
The testing result of amyloid beta protein 42 in 1 sample of table
The rate of recovery that 1 testing result of table can be seen that 42 testing result of amyloid beta protein in sample is 94~112 %, shows this
Invention can be applied to the detection of practical biological sample, as a result accurately and reliably.
Claims (4)
1. a kind of gold hydridization MoS2/Bi2S3The electrochemical luminescence sensor of the immobilized luminol detection amyloid beta protein 42 of nanometer rods,
Preparation step is as follows:
(1) using the glass-carbon electrode of polishing powder pretreatment 4 mm of diameter, ultrapure water is clean；
(2) by 7 μ L, 10 mg/mL luminol-gold hydridization MoS2/Bi2S3Nanometer rods solution drop coating is to electrode surface, room temperature preservation
To drying；
(3) 6 μ L of drop coating, 500 μ g/mL primary antibody Ab1Solution saves in glassy carbon electrode surface, 4 DEG C of refrigerators to drying, ultrapure water
Cleaning；
(4) bovine serum albumin(BSA) that 3 μ L mass fraction of drop coating is 1% is closed nonspecific activity site, is saved in 4 DEG C of refrigerators
To drying, ultrapure water cleaning；
(5) 42 drop coating of amyloid beta protein of 6 μ L various concentrations is saved in electrode surface, 4 DEG C of refrigerators to drying, it is ultrapure
Water cleaning；
(6) by 6 μ L, 1~7 mg/mL Ab2Poly-dopamine@curcumin-ZnO solution drop coating is in electrode surface, 4 DEG C of refrigerators
It saves to drying, ultrapure water cleaning obtains the electrochemical luminescence biosensor of detection amyloid beta protein 42.
2. a kind of golden hydridization MoS of the present invention2/Bi2S3The electrification of the immobilized luminol detection amyloid beta protein 42 of nanometer rods
Learn luminescence sensor, the luminol-gold hydridization MoS2/Bi2S3Nanometer rods solution, preparation step are as follows:
(1) MoS2/Bi2S3The preparation of nanometer rods
By 0.242 g Na2MoO4·2H2O and 0.765 g Bi (NO3)3·5H2O is dispersed in 60 mL ultrapure waters, is added 0.76
G thiocarbamide, 1 h of magnetic agitation；Above-mentioned solution is transferred in 100 mL reaction kettles, 220 DEG C of 24 h of reaction, centrifugation, is done washing
It is dry to obtain MoS2/Bi2S3Nanometer rods；
(2) luminol-gold hydridization MoS2/Bi2S3The preparation of nanometer rods
By the MoS of preparation2/Bi2S3Nanometer rods are dispersed in 50 mL ultrapure waters, 1 h of ultrasound；Then, by 2 mL, 1% HAuCl4With
5 mg PVP are added in above-mentioned solution, and after stirring 6 h, 2 mL, 50 mmol/L sodium citrate solution and a small amount of is added dropwise
NaBH4Solution reduction HAuCl4；After stirring 6 h, the unbonded gold nanoparticle of centrifugation removal obtains the MoS of golden hydridization2/
Bi2S3Nanometer rods；Then nanometer rods are dispersed in 5 mL ultrapure waters, 1~5 mL 5mmol/L luminol are added and is stirred overnight,
Pass through gold-NH2Luminol is incorporated in nano-bar material surface by key, and the unbonded luminol of centrifuge separation removal obtains Rumi
Nuo-gold hydridization MoS2/Bi2S3Nanometer rods.
3. a kind of golden hydridization MoS of the present invention2/Bi2S3The electrification of the immobilized luminol detection amyloid beta protein 42 of nanometer rods
Learn luminescence sensor, the Ab2Poly-dopamine@curcumin-ZnO, preparation step are as follows:
(1) preparation of curcumin-ZnO
5 mg curcumins are dispersed in 50 mL ultrapure waters, 90 DEG C are back to curcumin and are completely dissolved；Then, 50 mL are added
0.1 mol/L zinc nitrate solution, 90 DEG C of 1 h of reflux；When above-mentioned solution is cooled to room temperature, 5 mL 0.2 are added in ice-water bath
Mol/L KOH stirs 1 h, forms orange-yellow colloidal suspensions, with the unbonded turmeric of ultrapure water and acetone washing removal
Element, vacuum drying obtain curcumin-ZnO；
(2) Ab2Poly-dopamine@curcumin-ZnO
Curcumin-the ZnO of 20 mg preparation and 1~5 mg dopamine are dispersed in 30 mL ultrapure waters, 6 h are stirred, centrifugation is gone
Except unbonded dopamine；Said mixture is dispersed in 10 Tris-HCl(pH=8.5 mmol/L 10 mL) in solution, stir
Mix 6 h, the unbonded poly-dopamine of centrifugation removal；Then, 1~10 mg poly-dopamine@curcumin-ZnO is dispersed in 5 mL phosphorus
In hydrochlorate buffer solution (pH=7.4), 500 μ L, 500 μ g/mL secondary antibody Ab is added2, 6 h are stirred under 4 C；Later, 100 are added
1% bovine serum albumin(BSA) of μ L, closes non-specific sites, and centrifugation obtains Ab2Poly-dopamine@curcumin-ZnO, is dispersed 1
ML is stored in 4 DEG C of refrigerators stand-by in phosphate buffer solution (pH=7.4).
4. preparation method of the present invention preparation based on luminol-gold hydridization MoS2/Bi2S3Nanometer rods construct electrochemistry
Luminescence sensor, for the detection of amyloid beta protein 42, steps are as follows:
(1) using reference electrode-Ag/AgCl electrode, to electrode-platinum electrode, obtained electrochemical luminescence sensor as work
Make electrode, be connected in the magazine of chemiluminescence detector, electrochemical workstation and chemiluminescence detector are linked together,
The high pressure of photomultiplier tube is set as 600 V, and scanning voltage is set as -0.2~0.6 V；
(2) it using the PBS buffer solution of the mmol/L hydrogen peroxide Han 1 mmol/L~8, is detected by Electrochemiluminescince different
The electrochemical luminescence signals intensity that the amyloid beta protein 42 of concentration generates；
The PBS buffer solution, pH=6.5~8.5 are with 1/15 mol/L Na2HPO4With 1/15 mol/L KH2PO4Match
System；
(3) according to the linear relationship of resulting electrochemical luminescence intensity value and 42 log concentration of amyloid beta protein, it is bent to draw work
Line.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1359421A1 (en) * | 2002-05-03 | 2003-11-05 | Commissariat A L'energie Atomique | Method of detection of molecular recognition by electrochemiluminescence |
| CN103649295A (en) * | 2011-05-13 | 2014-03-19 | 加利福尼亚大学董事会 | Photothermal substrates for selective transfection of cells |
| KR20140056014A (en) * | 2012-10-29 | 2014-05-09 | 연세대학교 산학협력단 | Polydopamine-peptide-immobilized substrates for feeder-free maintenance and enhanced differentiation of stem cells and methods for preparing the substrates |
| CN108452376A (en) * | 2018-07-03 | 2018-08-28 | 南通大学 | A kind of poly-dopamine gelatine microsphere preparation method |
-
2019
- 2019-01-16 CN CN201910038073.2A patent/CN109507174B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1359421A1 (en) * | 2002-05-03 | 2003-11-05 | Commissariat A L'energie Atomique | Method of detection of molecular recognition by electrochemiluminescence |
| CN103649295A (en) * | 2011-05-13 | 2014-03-19 | 加利福尼亚大学董事会 | Photothermal substrates for selective transfection of cells |
| KR20140056014A (en) * | 2012-10-29 | 2014-05-09 | 연세대학교 산학협력단 | Polydopamine-peptide-immobilized substrates for feeder-free maintenance and enhanced differentiation of stem cells and methods for preparing the substrates |
| CN108452376A (en) * | 2018-07-03 | 2018-08-28 | 南通大学 | A kind of poly-dopamine gelatine microsphere preparation method |
Non-Patent Citations (1)
| Title |
|---|
| JIANG MENG等: "Advances in Detection Methods of β-Amyloid Protein", 《CHINESE JOURNAL OF ANALYTICAL CHEMISTRY》 * |
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