CN109709327A - Photoelectricity immunosensor based on water-soluble Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material - Google Patents
Photoelectricity immunosensor based on water-soluble Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material Download PDFInfo
- Publication number
- CN109709327A CN109709327A CN201811572333.6A CN201811572333A CN109709327A CN 109709327 A CN109709327 A CN 109709327A CN 201811572333 A CN201811572333 A CN 201811572333A CN 109709327 A CN109709327 A CN 109709327A
- Authority
- CN
- China
- Prior art keywords
- solution
- quantum dot
- titanium dioxide
- water
- dioxide nano
- 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.)
- Withdrawn
Links
Landscapes
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention belongs to tumor-marker analyte detection, optical electro-chemistry, immune sensing technical fields, specifically disclose a kind of water solubility Zn-Mg-Te quantum dot and its water solubility Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material preparation method, L-type glass-carbon electrode is modified based on water-soluble Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material, squamous cell anticancrin is loaded to construct photoelectricity immunosensor, based on immune response, and using white light as the specific detection of excitation light source realization squamous cell carcinoma antigen；Water-soluble Zn-Mg-Te quantum dot, titanium dioxide nano-rod and white light excitation light source are introduced squamous cell carcinoma photoelectricity immune sensing system by the present invention, prepared optical-electronic immunosensor have conveniently, simple, economy, strong applicability, it is easy to operate quickly, the advantages such as selectivity is good, high sensitivity, detection limit are low.
Description
Technical field
Present invention simultaneously relates to stigmata analyte detection, optical electro-chemistry, immune sensing technical fields, and in particular to Yi Zhongxin
Type water solubility Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material preparation, characterization and quantum dot and nano-silica
Change application of the energy transfer between titanium in squamous cell carcinoma photoelectricity immunosensor.
Background technique
It is shown according to global cancer statistical data, cervical carcinoma can all lead to about 250,000 women dies every year, wherein absolutely mostly
Several cervical carcinomas is to belong to squamous cell type；Squamous cell carcinoma antigen (SCCA) is a kind of TA-4 hypotype glycoprotein, is confirmed as
Clinically for detecting a kind of tumor markers of cervical carcinoma, the raising of internal SCCA content also with lung cancer, head and neck cancer, vagina
Cancer, liver cancer etc. are related, in-vivo content extremely low (0 ~ 1.5ng mL of the SCCA in normal person-1), document is it has been reported that for examining at present
The method for surveying SCCA mainly has enzyme-linked immunization, chemoluminescence method etc., but these methods have sensitivity low or emit the strong of light
Spend the problems such as high to the dependency degree of environment.
Optical electro-chemistry sensor is the sensing device that determinand is detected using the photoelectric characteristic of substance, is hair recent years
A kind of analysis method that exhibition is got up；Optical electro-chemistry is as a kind of analysis means, its detection process and electrogenerated chemiluminescence is just
On the contrary；Due to using various forms of excitations and detection signal, thus its background signal is lower, after some time it is possible to reach sends out with electroluminescent chemistry
The comparable high sensitivity of light；Also, the instrument of optical electro-chemistry is fairly simple, is easy micromation；Due to using Electrochemical Detection, together
Optical detecting method is compared, and equipment is more inexpensive；In fact, being based on when carrying out same substance detection using same design
The method of optical electro-chemistry sensor also shows better detection performance (such as lower detection limit) than the method based on electrochemistry；
Based on the above advantage, optical electro-chemistry sensor has broad application prospects in terms of determination of biomolecule；But optical electro-chemistry passes
Sense technology is still in infancy at present, primarily focuses on the utilization of novel optoelectronic materials, the building of novel detection method, detection
The diversification of substance.
Quantum dot is a kind of important photoelectric semiconductor material, is widely used in optical electro-chemistry sensing Study of An, but single amount
Son point material causes photoelectric conversion efficiency lower due to the compound of internal electron hole pair；Therefore, a suitable method is found,
The photoelectric conversion efficiency for effectively promoting photoelectric material is very important；With the progress of new material technology, nano material
Importance is increasingly prominent, it is filled with new strength for optical electro-chemistry sensor；Especially nanocomposite compares homogenous material
With higher photoelectric conversion efficiency；In recent years, the research of nanocomposite obtains extensive concern.
Titanium dioxide nano material is due to its excellent biocompatibility, excellent chemical stability, bigger serface, machine
The advantages that tool performance is good, simple synthetic method has attracted extensive concern, especially one-dimensional titanium dioxide nanostructure, such as dioxy
Change titanium nanometer rods, is widely used in the preparation of optical electro-chemistry and electrochemical immunosensor.
Summary of the invention
In order to overcome above-mentioned the deficiencies in the prior art, it is an object of the invention to:
First, a kind of preparation method of water solubility Zn-Mg-Te quantum dot is provided；
Second, the water-soluble Zn-Mg-Te quantum dot/titanium dioxide nano-rod for providing a kind of novel photoelectric high conversion efficiency is compound
The preparation method of material；
Third provides a kind of based on above-mentioned new type water-solubility Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material modification L
The photoelectricity immunosensor of type glass-carbon electrode；
4th, application of the above-mentioned photoelectricity immunosensor in measurement squamous cell carcinoma is provided.
A kind of preparation method of water solubility Zn-Mg-Te quantum dot, comprises the following steps that
(1) preparation of NaHTe solution: under nitrogen protection, by 80mg NaBH4, 10ml water sequentially adds in 25ml three-necked flask,
Ice bath reaction, is added 127mg tellurium powder, reacts 4h to black tellurium powder fully reacting and obtains colorless and transparent NaHTe solution；
(2) under nitrogen protection, 60mL 0.002mol/L ZnCl is sequentially added into three neck round bottom2Solution, 20mL
0.016mol/L N-acetyl-L-cysteine solution and 20mL secondary distilled water are uniformly mixed, obtain mixed liquor；Use 1mol/L
The pH value that NaOH solution adjusts mixed liquor is 11-11.5；0.2-2mL 0.01mol/L MgCl is added2Solution is protected from light
0.5h；Then the NaHTe solution of 3mL step (1) is added, 100 DEG C of reaction 4-5h obtain Zn-Mg-Te quantum dot solution, cooling
To room temperature, it is centrifuged with the super filter tube of 10kD, obtains water-soluble Zn-Mg-Te quantum dot solution, 4 DEG C save backup.
A kind of preparation method of water solubility Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material, including step is such as
Under:
The preparation of S1, titanium dioxide nano-rod: under ice-water bath, 1.8ml titanium tetrachloride is slowly added into the ultrapure water containing 19ml
In beaker, strong stirring 10min obtains white suspension, then 1.8ml chloroform is added in this mixed liquor, continues to stir 10min,
Mixed liquor is transferred in the autoclave of polytetrafluoroethyllining lining, 180 DEG C of reaction 6h are cooled to room temperature, white is collected by centrifugation
Precipitating, is washed with deionized water and dehydrated alcohol to pH7.0, then 60 DEG C of vacuum drying 12h, store for future use at room temperature；
S2, titanium dioxide nano-rod amination: the 100mg titanium dioxide nano-rod that step S1 is prepared is placed in second containing 20ml
In the mixed liquor of alcohol, the ammonia spirit that 1ml mass concentration is 28% and 4ml3 aminopropyl triethoxysilane, stir at room temperature
Overnight, next day centrifuge separation discards supernatant liquid and stores for future use at room temperature after the washing of gained white depositions, drying；
S3, take 100 μ L be added in 1ml Zn-Mg-Te quantum dot solution contain 1-(3- dimethylamino-propyl) -3- ethyl carbon two
The mixed solution of inferior amine salt hydrochlorate and n-hydroxysuccinimide, is uniformly mixed, and adds amino made from 5mg step S2
Oxide/titanium dioxide nanometer rods, stir 4-6h at room temperature, with secondary distilled water centrifuge washing 3 times, are finally diluted with secondary distilled water
To 1 mL, water-soluble Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material, abbreviation Zn-Mg-Te/TiO are obtained2Composite wood
Material；
The mixing containing 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and n-hydroxysuccinimide is molten
Liquid is the mixed aqueous solution of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and n-hydroxysuccinimide, is mixed
1-(3- dimethylamino-propyl -3- ethyl-carbodiimide hydrochloride concentration is 10 mg/mL in Heshui solution, and N- hydroxysuccinimidyl acyl is sub-
Amine concentration is 20 mg/mL.
A kind of squamous cell carcinoma photoelectricity based on water-soluble Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material is exempted from
The preparation method of epidemic disease sensor, its step are as follows:
(1) pretreatment of glassy carbon electrode: the bare glassy carbon electrode of L-type, 3 mm of diameter, with 0.3 μm of Al of granularity2O3Suspension polishing,
Distilled water flushing is clean, then successively in HNO3, ethyl alcohol, be cleaned by ultrasonic in secondary distilled water, dry spare；
(2) Zn-Mg-Te/TiO of the 6 above-mentioned preparations of μ L is taken2Clean glass-carbon electrode table obtained by composite material drop coating to step (1)
Face is dried at room temperature, is cleaned with the PBS buffer solution of 0.01 pH=7.4 mol/L, is obtained Zn-Mg-Te/TiO2/ GCE modification
Electrode；
Take the squamous cell anticancrin anti-SCCA solution drop coating of 6 μ L, 10 μ g/mL to Zn-Mg-Te/TiO again2/ GCE modification electricity
Pole surface is dried at room temperature, is cleaned loose anti-SCCA with the PBS buffer solution of 0.01 pH=7.4 mol/L, is obtained
anti-SCCA/Zn-Mg-Te/TiO2/ GCE modified electrode；
Again by anti-SCCA/Zn-Mg-Te/TiO2/ GCE modified electrode is crosslinked 5 min in glutaraldehyde steam, with 0.01
The PBS buffer solution of pH=7.4 mol/L is cleaned, and is dried；0.5 h is impregnated, in 0.25wt% bovine serum albumin solution with envelope
Electrode surface nonspecific binding site that may be present is closed, is washed after taking-up with the PBS buffer solution of 0.01 pH=7.4 mol/L
Only, it dries, obtains squamous cell carcinoma photoelectricity immunosensor, be placed in 4 DEG C of refrigerators and save for use.
Using white light as light source, to above-mentioned in the 0.1mol/L phosphate buffer (pH 7.4) of 0.25mol/L ascorbic acid
Squamous cell carcinoma photoelectricity immunosensor carry out Photoelectronic property test, and be used for squamous cell carcinoma antigen measurement, have compared with
Good sensitivity, accuracy, precision and selectivity.
At 35 DEG C, the squamous cell carcinoma of various concentration is respectively placed in above-mentioned squamous cell carcinoma optical-electronic immunosensor
In SCCA antigenic solution, concentration be respectively as follows: 1pg/mL, 5pg/mL, 10pg/mL, 100 pg/mL, 1ng/mL, 10ng/mL,
50ng/mL is incubated for 30min, is then cleaned with 0.01mol/L PBS buffer solution (pH7.4), must combine the squama of SCCA antigen
Shape cell cancer optical-electronic immunosensor, 4 DEG C of storages are ready for use on Photoelectronic property test.
Compared with prior art, the advantages and beneficial effects of the present invention are:
(1) present invention exempts from using a kind of new type water-solubility Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material as photoelectricity
The sensitive material of epidemic disease sensor, the material have strong and stable photoelectric current, can be realized to squamous under white light source excitation
The highly sensitive detection of cell cancer, detection are limited to 1.89pg/mL(S/N=3).
(2) water-soluble Zn-Mg-Te ternary quantum dots are introduced optical electro-chemistry and sense system by the present invention, which is free of
The heavy metal ion such as Cd, Hg, good biocompatibility, Wavelength tunable have stronger fluorescence intensity and stability.
(3) present invention is using Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material as photoelectric sensor, through solid
Squamous cell carcinoma photoelectricity immunosensor, titanium dioxide nano-rod good conductivity, biocompatibility are prepared after carrying anti-SCCA
Good, large specific surface area, helps to maintain antibody bioactive, and improve detection sensitivity.
(4) operating process of photoelectricity immunosensor of the invention measurement squamous cell carcinoma is easy, does not need special experiment
Condition, instrument requirements are simple, can be used for the clinical examination of the patients such as cervical carcinoma, lung cancer, head and neck cancer, carcinoma of vagina, liver cancer, in medical treatment
Field is with good application prospect.
Detailed description of the invention
Fig. 1 is a kind of fluorescence spectra of water solubility Zn-Mg-Te quantum dot prepared by embodiment 1.
Fig. 2 is a kind of transmission electron microscope picture of water solubility Zn-Mg-Te quantum dot of 1 scheme one of embodiment preparation.
Fig. 3 is a kind of water solubility Zn-Mg-Te quantum of N-acetyl-L-cysteine (a) and the preparation of 1 scheme one of embodiment
The infrared spectrogram of point (b).
Fig. 4 is the transmission electron microscope picture of titanium dioxide nano-rod prepared by embodiment 2.
Fig. 5 is Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material transmission electron microscope picture prepared by embodiment 3.
Fig. 6 is Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material scanning electron microscope (SEM) photograph prepared by embodiment 3.
Fig. 7 is Zn-Mg-Te/GCE(a), Zn-Mg-Te/TiO2/ GCE(b) photo-current intensity comparison diagram.
Fig. 8 is SCCA photoelectricity immunosensor preparation process schematic diagram of the invention.
Fig. 9 is naked GCE electrode (curve a), Zn-Mg-Te/TiO prepared by the present invention2/ GCE curve (curve b),
anti-SCCA/Zn-Mg-Te/TiO2/ GCE(curve c), anti-SCCA(BSA)/Zn-Mg-Te/TiO2/ GCE(curve d),
SCCA/ anti-SCCA(BSA)/Zn-Mg-Te/TiO2The AC impedance figure of/GCE(curve e).
Figure 10 is Zn-Mg-Te/TiO prepared by the present invention2/ GCE(curve a), anti-SCCA/Zn-Mg-Te/TiO2/
GCE(curve b), anti-SCCA(BSA)/Zn-Mg-Te/TiO2/ GCE(curve c), SCCA/ anti-SCCA(BSA)/Zn-
Mg-Te/TiO2/ GCE(curve d) photocurrent-time response curve.
Figure 11 is the calibration curve that immunosensor of the invention detects SCCA.
Figure 12 is immunosensor of the invention in noiseless object (a) and photocurrent response in the presence of having chaff interferent (b)
Figure.
Specific embodiment
Combined with specific embodiments below, the present invention is furture elucidated；It should be understood that these embodiments are merely to illustrate the present invention
Rather than the claimed range of the limitation present invention；It will further be understood that read the content lectured of the present invention it
Afterwards, those skilled in the art can make various changes or modification to the present invention, and such equivalent forms are equally fallen within appended by the application
Claims limited range.
White light used is that 5W highlights external white light source in following embodiment, is purchased from " light concept car decorations shop " Taobao
Shop, electrode material L-type glass-carbon electrode are bought in Tianjin Grolsch Rui Lian Science and Technology Ltd., glass carbon di 3mm；Anti-SCCA
McAb (labeling) (L1C01402), squamous cell carcinoma SCCA antigen (L2C01501) are purchased from Shanghai and lead damp biotechnology
Co., Ltd is diluted to required concentration with 0.01mol/LPBS buffer (pH=7.4)；It with water is second distillation in embodiment
Water.
Embodiment 1
A kind of preparation method of water solubility Zn-Mg-Te quantum dot, comprises the following steps that
The preparation of NaHTe solution: under nitrogen protection, by 80mg NaBH4,10ml water is sequentially added in 25ml three-necked flask, ice bath
127mg tellurium powder is added in reaction, reacts 4h to black tellurium powder fully reacting and obtains colorless and transparent NaHTe solution, spare；
Scheme one: under nitrogen protection, 60mL 0.002mol/L ZnCl is sequentially added into three neck round bottom2Solution, 20mL
0.016mol/L N-acetyl-L-cysteine solution and 20mL secondary distilled water are uniformly mixed, obtain mixed liquor；Use 1mol/L
The pH value that NaOH solution adjusts mixed liquor is 11.5；0.5mL 0.01mol/L MgCl is added2Solution is protected from light 0.5h；So
The freshly prepd NaHTe solution of 3mL is added afterwards, 100 DEG C of reaction 5h, obtaining fluorescence emission wavelengths is 580nm, as shown in Fig. 1 curve e
Zn-Mg-Te quantum dot solution；It is cooled to room temperature, with the super filter tube 8000rpm refrigerated centrifuge 10min of 10kD, obtains 100 μ L
New type water-solubility ternary Zn-Mg-Te quantum dot, 4 DEG C save backup；And this wavelength quantum dot is used for subsequent optical Electro-immune sensing
Process；Its pattern is characterized with transmission electron microscope, and TEM figure is as shown in Figure 2；As can be observed from Figure, which is in ball
Shape, favorable dispersibility, size is in 4-5nm or so.
Scheme two: under nitrogen protection, 60mL 0.002mol/L ZnCl is sequentially added into three neck round bottom2Solution,
20mL 0.016mol/L N-acetyl-L-cysteine solution and 20mL secondary distilled water are uniformly mixed, obtain mixed liquor；With
The pH value that 1mol/L NaOH solution adjusts mixed liquor is 11.0；0.2mL 0.01mol/L MgCl is added2Solution is protected from light
0.5h；Then the freshly prepd NaHTe solution of 3mL is added, 100 DEG C of reaction 4h, obtaining fluorescence emission wavelengths is 520nm, such as Fig. 1 bent
Zn-Mg-Te quantum dot solution shown in line a；It is cooled to room temperature, with the super filter tube 8000rpm refrigerated centrifuge 10min of 10kD, obtains
To 100 μ L new type water-solubility ternary Zn-Mg-Te quantum dots, 4 DEG C are saved backup.
Scheme three: under nitrogen protection, 60mL 0.002mol/L ZnCl is sequentially added into three neck round bottom2Solution,
20mL 0.016mol/L N-acetyl-L-cysteine solution and 20mL secondary distilled water are uniformly mixed, obtain mixed liquor；With
The pH value that 1mol/L NaOH solution adjusts mixed liquor is 11.5；1mL 0.01mol/L MgCl is added2Solution is protected from light
0.5h；Then the freshly prepd NaHTe solution of 3mL is added, 100 DEG C of reaction 4h, obtaining fluorescence emission wavelengths is 530nm, such as Fig. 1 bent
Zn-Mg-Te quantum dot solution shown in line b；It is cooled to room temperature, with the super filter tube 8000rpm refrigerated centrifuge 10min of 10kD, obtains
To 100 μ L new type water-solubility ternary Zn-Mg-Te quantum dots, 4 DEG C are saved backup.
Scheme four: under nitrogen protection, 60mL 0.002mol/L ZnCl is sequentially added into three neck round bottom2Solution,
20mL 0.016mol/L N-acetyl-L-cysteine solution and 20mL secondary distilled water are uniformly mixed, obtain mixed liquor；With
The pH value that 1mol/L NaOH solution adjusts mixed liquor is 11.5；1.5mL 0.01mol/L MgCl is added2Solution is protected from light
0.5h；Then the freshly prepd NaHTe solution of 3mL is added, 100 DEG C of reaction 4h, obtaining fluorescence emission wavelengths is 550nm, such as Fig. 1 bent
Zn-Mg-Te quantum dot solution shown in line c；It is cooled to room temperature, with the super filter tube 8000rpm refrigerated centrifuge 10min of 10kD, obtains
To 100 μ L new type water-solubility ternary Zn-Mg-Te quantum dots, 4 DEG C are saved backup.
Scheme five: under nitrogen protection, 60mL 0.002mol/L ZnCl is sequentially added into three neck round bottom2Solution,
20mL 0.016mol/L N-acetyl-L-cysteine solution and 20mL secondary distilled water are uniformly mixed, obtain mixed liquor；With
The pH value that 1mol/L NaOH solution adjusts mixed liquor is 11.5；2mL 0.01mol/L MgCl is added2Solution is protected from light
0.5h；Then the freshly prepd NaHTe solution of 3mL is added, 100 DEG C of reaction 5h, obtaining fluorescence emission wavelengths is 560nm, such as Fig. 1 bent
Zn-Mg-Te quantum dot solution shown in line d；It is cooled to room temperature, with the super filter tube 8000rpm refrigerated centrifuge 10min of 10kD, obtains
To 100 μ L new type water-solubility ternary Zn-Mg-Te quantum dots, 4 DEG C are saved backup.
Fig. 3 be N-acetyl-L-cysteine (the Zn-Mg-Te quantum dot that curve a) and 1 scheme one of embodiment obtain
Infrared spectrogram (curve b)；By curve a it is found that 2547.48cm-1For the characteristic absorption 1717.16cm of sulfydryl-1 For C=O bond
Characteristic absorption, 1586.61cm-1For the characteristic absorption peak of the in-plane bending vibration of-NH in amide, 2900.71cm-1With
2808.78cm-1Two characteristic absorption peaks show containing methylene；It will be evident that the key band of sulfydryl from curve b
2547.48cm-1It disappears on the spectrogram of quantum dot after modification, in addition, methylene characteristic absorption still remains, it follows that mercapto
Base and quantum dot may be bonded by Zn-S to be closed, and shows that successfully Zn-Mg-Te quantum is arrived in modification to N-acetyl-L-cysteine
Point surface.
Embodiment 2
A kind of preparation method of water solubility Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material, comprises the following steps that
The preparation of S1, titanium dioxide nano-rod: under ice-water bath, 1.8ml titanium tetrachloride is slowly added into the ultrapure water containing 19ml
In beaker, strong stirring 10min obtains white suspension, then 1.8ml chloroform is added in this mixed liquor, continues to stir 10min,
Mixed liquor is transferred in the autoclave of polytetrafluoroethyllining lining, 180 DEG C of reaction 6h are cooled to room temperature, white is collected by centrifugation
Precipitating, is washed with deionized water and dehydrated alcohol to pH7.0, then 60 DEG C of vacuum drying 12h, store for future use at room temperature；
S2, titanium dioxide nano-rod amination: the 100mg titanium dioxide nano-rod that step S1 is prepared is placed in second containing 20ml
In the mixed liquor of alcohol, the ammonia spirit that 1ml mass concentration is 28% and 4ml3 aminopropyl triethoxysilane, stir at room temperature
Overnight, next day centrifuge separation discards supernatant liquid and stores for future use at room temperature after the washing of gained white depositions, drying；
S3, take 100 μ L be added in 1ml Zn-Mg-Te quantum dot solution contain 1-(3- dimethylamino-propyl) -3- ethyl carbon two
The mixed solution of inferior amine salt hydrochlorate and n-hydroxysuccinimide, is uniformly mixed, and adds amino made from 5mg step S2
Oxide/titanium dioxide nanometer rods, stir 4h at room temperature, with secondary distilled water centrifuge washing 3 times, are finally diluted to 1 with secondary distilled water
ML obtains water-soluble Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material, abbreviation Zn-Mg-Te/TiO2Composite material；
The mixing containing 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and n-hydroxysuccinimide is molten
Liquid is the mixed aqueous solution of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and n-hydroxysuccinimide, is mixed
1-(3- dimethylamino-propyl -3- ethyl-carbodiimide hydrochloride concentration is 10 mg/mL in Heshui solution, and N- hydroxysuccinimidyl acyl is sub-
Amine concentration is 20 mg/mL.
By transmission electron microscope as shown in figure 4, all titanium dioxide nano-rods are cylindrical structure, average diameter is about
20nm, average length are about 120nm, this facilitates the load of biomolecule.
Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material transmission electron microscope picture, scanning electron microscope (SEM) photograph are respectively as schemed
5, shown in Fig. 6, You Tuzhong is the results show that titanium dioxide nano-rod surface has been capped a large amount of quantum dot particle, surface Zn-
Mg-Te quantum dot is by covalent bond and has successfully been assembled into golden stick surface.
In order to prove titanium dioxide nano-rod to the photoelectricity enhancement effect of Zn-Mg-Te quantum dot, identical item in experimental study
Under part, we are to Zn-Mg-Te quantum dot, Zn-Mg-Te/TiO2The photoelectric current that composite material modified electrode generates has carried out pair
Than；Under identical conditions, we load ZnMgTe quantum dot composite material to Zn-Mg-Te quantum dot and titanium dioxide nano-rod
The photoelectric current generated on glass-carbon electrode is compared, as shown in Figure 7；It is multiple that titanium dioxide nano-rod loads ZnMgTe quantum dot
The photoelectric current of about 3.4 μ A is generated on condensation material (b) electrode, in contrast, the individually electrode of ZnMgTe quantum dot (a) modification
About 1.6 μ A of photoelectric current, it is possible thereby to prove that titanium dioxide nano-rod has significant humidification to photoelectric current；Titanium dioxide
Nanometer rods are as follows to photoelectric current enhancing mechanism: firstly, the attached of quantum dot can be improved in the big specific surface area of titanium dioxide nano-rod
Amount, thus more quantum dot can be attached to its surface to capture more light；Secondly, titanium dioxide nano-rod can be used as
Electron acceptor is to accelerate the transmitting of electronics and the electron-hole of excitation state quantum dot can be hindered compound simultaneously.
About in case study on implementation 2, titanium dioxide nano-rod in Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material
It, can be compound by investigating Zn-Mg-Te quantum dot/titanium dioxide nano-rod of different proportion with the best preparation ratio of quantum dot
Photocurrent-time of the material modified electrode in the 0.1mol/LPBS buffer solution (pH=7.4) of 0.25mol/L ascorbic acid
Response curve determines；With the increase of quantum dot ratio, the photo-current intensity of corresponding composite material is gradually increased, when two
When the ratio between amount of substance of TiOx nano rod composite material and quantum dot is 1:80, the photoelectric current of corresponding composite material
Reach maximum, continues growing the photo-current intensity reduction of its corresponding composite material of quantum dot ratio, this may be due to excessive
The exchange conduction of the increase of quantum dot, electronics is suppressed.
Embodiment 3
As shown in figure 8, a kind of squamous cell carcinoma based on water-soluble Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material
The preparation method of photoelectricity immunosensor, its step are as follows:
(1) bare glassy carbon electrode (diameter 3mm) of type is polished with 0.3 μm of Al2O3 suspension, and distilled water flushing is clean, then according to
It is secondary in HNO3Ultrasound is clear in (being formed after being mixed in equal volume by 65wt% concentrated nitric acid with water), ethyl alcohol (50v/v%), secondary distilled water
It washes, dries spare；
(2) SCCA photoelectricity immunosensor is constructed:
The Zn-Mg-Te/TiO for taking 6 μ L embodiments 2 to prepare2Clean glass-carbon electrode table obtained by composite material drop coating to step (1)
Face is dried at room temperature, is cleaned with the PBS buffer solution of 0.01 pH=7.4 mol/L, is obtained Zn-Mg-Te/TiO2/ GCE modification
Electrode；
Take the squamous cell anticancrin anti-SCCA solution drop coating of 6 μ L, 10 μ g/mL to Zn-Mg-Te/TiO again2/ GCE modification electricity
Pole surface is dried at room temperature, is cleaned loose anti-SCCA with the PBS buffer solution of 0.01 pH=7.4 mol/L, is obtained
anti-SCCA/Zn-Mg-Te/TiO2/ GCE modified electrode；
Again by anti-SCCA/Zn-Mg-Te/TiO2/ GCE modified electrode is crosslinked 5 min in glutaraldehyde steam, with 0.01
The PBS buffer solution of pH=7.4 mol/L is cleaned, and is dried；0.5 h is impregnated, in 0.25wt% bovine serum albumin solution with envelope
Electrode surface nonspecific binding site that may be present is closed, is washed after taking-up with the PBS buffer solution of 0.01 pH=7.4 mol/L
Only, dry, obtain anti-SCCA(BSA)/Zn-Mg-Te/TiO2/ GCE modified electrode is to get squamous cell carcinoma photoelectricity immune sensing
Device is placed in 4 DEG C of refrigerators and saves for use；
By anti-SCCA(BSA)/Zn-Mg-Te/TiO2/ GCE modified electrode is respectively placed in the squamous cell carcinoma of various concentration
In SCCA antigenic solution, concentration be respectively as follows: 1pg/mL, 5pg/mL, 10pg/mL, 100 pg/mL, 1ng/mL, 10ng/mL,
Then 50ng/mL, 35 DEG C of incubation 30min are cleaned with 0.01mol/LPBS buffer (pH7.4), obtain SCCA/ anti-SCCA
(BSA)/Zn-Mg-Te/TiO2/ GCE modified electrode is to get the squamous cell carcinoma optical-electronic immune sensing for combining SCCA antigen
Device, 4 DEG C of storages are ready for use on Photoelectronic property test.
(3) self assembly optical electro-chemistry test macro: using 5W white light as excitation light source, guiding through light to electrode surface,
Use three-electrode system test light electric current: using the L-type glass-carbon electrode of 3mm as working electrode, Hg/Hg2Cl2Electrode (saturation
KCl it) is used as reference electrode, platinum electrode is used as to electrode, and photoelectric current is by CHI660E electrochemical workstation (Shanghai Chen Hua instrument
Company) measurement；Photoelectric current is in constant potential (0V vs Hg/Hg2Cl2), the 0.1mol/LPBS of 0.25mol/L ascorbic acid buffering
It is carried out in solution (pH=7.0).
About in step (2), the best incubation time of SCCA antigen can be by investigating the photocurrent response of sensor and incubating
Educate the relation curve of time to determine: within the scope of 10~40min, as incubation time extends, immunosensor identifies SCCA
The photocurrent response difference of front and back quicklys increase, and tends towards stability after 30min, shows that immune combine of Ag-Ab tends to be saturated, because
This, the present invention selects 30min for best incubation time.
About in step (2), the best incubation temperature of SCCA antigen can be by investigating the photocurrent response of sensor and incubating
The relation curve of time is educated to determine: within the scope of 20~35 DEG C, being increased with incubation temperature, and immunosensor identifies before and after SCCA
Photocurrent response difference quickly increase, 35 DEG C of whens, reach maximum value；After temperature is higher than 35 DEG C, protein active will affect,
Its recognition capability is reduced, photocurrent response difference reduces；Therefore, the present invention select 35 DEG C for best incubation temperature.
About in step (3), the concentration of ascorbic acid is by same sensor in various concentration ascorbic acid
Photocurrent-time response curve in 0.1mol/L phosphate buffer solution (pH 7.0) determines；From 0 mol/L~0.25mol/
L, as ascorbic acid concentrations gradually increase, photoelectric current is also gradually increased；It is further increased with ascorbic acid concentrations, photoelectric current subtracts
It is small；This is because concentration is too big, the absorbance of ascorbic acid in the solution increases, so as to cause the light intensity for being irradiated to electrode surface
Degree declines and the launching efficiency of quantum dot is caused to reduce；Accordingly, it is considered to the sensibility of electrode response Ascorbic Acid concentration is arrived, this
Invention selects optium concentration of the 0.25mol/L as ascorbic acid.
The above-mentioned optimal parameter condition decided also is selected in following photoelectrochemical behaviour test.
Embodiment 4
As shown in figure 9, electrochemical alternate impedance spectrum (EIS) characterization is carried out to modified electrode obtained by each step of embodiment 3,
EIS is one of the effective tool for exploring chemically modified electrode interfacial property;Its figure is made of low frequency range and high frequency region two parts,
In low frequency range correspond to diffusion control zone, and the high frequency region of semi-circular portions then corresponds to dynamics Controlling area, semicircle it is straight
Diameter size reflects electrode surface charge transfer resistance size；In 5.0mmol/L K3[Fe(CN)6]/K4[Fe(CN)6] (1:1)+
AC impedance characterization 0.1mol/LPBS(pH=7.0) is carried out in+0.1mol/L KCl solution；It is all kinds of in sensor preparation process
The Nyquist curve of modified electrode is as shown in Figure 8, it can be seen that (ac impedance spectroscopy of curve a) is in high frequency section for bare electrode
Semicircle very little；Work as Zn-Mg-Te/TiO2Composite material is modified to electrode surface, and half circular diameter of high frequency section increases
(curve b), impedance increase；Curve c is anti-SCCA/Zn-Mg-Te/TiO2The ac impedance spectroscopy at/GCE modified electrode interface
Figure, since antibody is detrimental to the large biological molecule of electron transmission, can hinder the electron transmission at interface, impedance significantly increases；
Behind nonspecific activity site that may be present with BSA enclosed-electrode surface, half circular diameter of ac impedance spectroscopy is greatly increased
(curve d)；After immunosensor is in conjunction with SCCA antigenic specificity, antigen-antibody complex covers electrode surface, interface
Charge-transfer resistance further increases (curve e)；The above result shows that the present invention after gradually modifying, successfully prepares SCCA
Photoelectricity immunosensor.
As shown in Figure 10, work as Zn-Mg-Te/TiO2/ GCE modified electrode (a) surface combine anti-SCCA antibody (b) and
BSA(c after) closing nonactive site, photoelectric current is reduced, this may be the inhibition due to albumen to electron transmission, makes molten
Electron donor ascorbic acid is difficult to diffuse to electrode surface in liquid, so as to cause the reduction of photo-current intensity；Work as immunosensor
After in conjunction with SCCA antigentic specificity (d), observe that photo-current intensity further decreases；Show modification step and immunoassays
Success carries out.
As shown in figure 11, the photoelectricity immunosensor in embodiment 3 has been investigated for various concentration with photocurrent-time method
The photocurrent response situation of SCCA antigen, when the concentration of SCCA is between 1pg/mL-50ng/mL, before and after bonding SCCA antigen
Photoelectric current difference be bonded the logarithm of the photoelectric current ratio before antigen and SCCA antigen concentration into good linear relationship, linear side
Journey are as follows: I=0.1105logcSCCA(ng/mL)+0.4325(R2=0.994), detection is limited to 1.89pg/mL(S/N=3)；This table
It is bright to be based on water solubility Zn-Mg-Te quantum dot and TiO2The photoelectricity immunosensor of energy transfer can be used for SCCA between nanometer rods
The highly sensitive detection of antigen.
Embodiment 5
Measure SCCA photoelectricity immunosensor (i.e. anti-SCCA(BSA)/Zn-Mg-Te/TiO obtained by embodiment 32/GCE
Modified electrode) selectivity, interference free performance be measure electrochemical sensor one of the important indicator of practicability；Prepare 5 parts
SCCA antigenic solution :(a) antigen containing 0.1ng/mLSCCA；(b) the human seralbumin egg of antigen containing 0.1ng/mLSCCA and 50ng/mL
White (HAS)；(c) alpha-fetoprotein (AFP) of antigen containing 0.1ng/mLSCCA and 50ng/mL；(d) antigen containing 0.1ng/mLSCCA and
The immunoglobulin (IgG) of 50ng/mL；(e) glycine (Gly) of antigen containing 0.1ng/mLSCCA and 50ng/mL；In best item
It under part, is measured after SCCA photoelectricity immunosensor is incubated for 30min in 5 parts of antigenic solutions respectively, as a result such as Figure 12,
As seen from the figure, the deviation of the current-responsive difference of (b)-(e) and (a) is respectively less than 5%, illustrates that the above substance does not interfere SCCA anti-
Former measurement, the i.e. immunosensor have good selectivity.
The above, only of the invention illustrates embodiment, not to the present invention in any form with substantial limitation,
It should be pointed out that for those skilled in the art, under the premise of not departing from the method for the present invention, that makes several changes
It also should be regarded as protection scope of the present invention into supplement；All those skilled in the art, do not depart from spirit of that invention and
In the case where range, using the equivalent variations of a little change, modification and differentiation that disclosed above technology contents are made, it is
Equivalent embodiment of the invention；Meanwhile any equivalent variations that all substantial technologicals according to the present invention do above-described embodiment
Change, modification and differentiation, still fall within protection scope of the present invention.
Claims (4)
1. a kind of preparation method of water solubility Zn-Mg-Te quantum dot, which is characterized in that comprise the following steps that
(1) preparation of NaHTe solution: under nitrogen protection, by 80mgNaBH4, 10ml water sequentially adds in 25ml three-necked flask, ice
Bath reaction, is added 127mg tellurium powder, reacts 4h to black tellurium powder fully reacting and obtains colorless and transparent NaHTe solution；
(2) under nitrogen protection, 60mL 0.002mol/L ZnCl is sequentially added into three neck round bottom2Solution, 20mL
0.016mol/L N-acetyl-L-cysteine solution and 20mL secondary distilled water are uniformly mixed, obtain mixed liquor；Use 1mol/L
The pH value that NaOH solution adjusts mixed liquor is 11-11.5；0.2-2mL 0.01mol/L MgCl is added2Solution is protected from light
0.5h；Then the NaHTe solution of 3mL step (1) is added, 100 DEG C of reaction 4-5h obtain Zn-Mg-Te quantum dot solution, cooling
To room temperature, it is centrifuged with the super filter tube of 10kD, obtains water-soluble Zn-Mg-Te quantum dot solution, 4 DEG C save backup.
2. a kind of preparation method of water solubility Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material, which is characterized in that packet
Include that steps are as follows:
The preparation of S1, titanium dioxide nano-rod: under ice-water bath, 1.8ml titanium tetrachloride is slowly added into the ultrapure water containing 19ml
In beaker, strong stirring 10min obtains white suspension, then 1.8ml chloroform is added in this mixed liquor, continues to stir 10min,
Mixed liquor is transferred in the autoclave of polytetrafluoroethyllining lining, 180 DEG C of reaction 6h are cooled to room temperature, white is collected by centrifugation
Precipitating, is washed with deionized water and dehydrated alcohol to pH7.0, then 60 DEG C of vacuum drying 12h, store for future use at room temperature；
S2, titanium dioxide nano-rod amination: the 100mg titanium dioxide nano-rod that step S1 is prepared is placed in second containing 20ml
It is stirred at room temperature in the mixed liquor of alcohol, the ammonia spirit that 1ml mass concentration is 28% and 4ml3 aminopropyl triethoxysilane
Night, next day centrifuge separation discard supernatant liquid and store for future use at room temperature after the washing of gained white depositions, drying；
S3, take 100 μ L be added in 1ml Zn-Mg-Te quantum dot solution contain 1-(3- dimethylamino-propyl) -3- ethyl carbon two
The mixed solution of inferior amine salt hydrochlorate and n-hydroxysuccinimide, is uniformly mixed, and adds amino made from 5mg step S2
Oxide/titanium dioxide nanometer rods, stir 4-6h at room temperature, with secondary distilled water centrifuge washing 3 times, are finally diluted with secondary distilled water
To 1 mL, water-soluble Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material, abbreviation Zn-Mg-Te/TiO are obtained2Composite wood
Material；
The mixing containing 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and n-hydroxysuccinimide is molten
Liquid is the mixed aqueous solution of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and n-hydroxysuccinimide, is mixed
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride concentration is 10 mg/mL, N- hydroxysuccinimidyl acyl in Heshui solution
Imines concentration is 20 mg/mL.
3. a kind of squamous cell carcinoma photoelectricity based on water-soluble Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material is immune
The preparation method of sensor, its step are as follows:
(1) pretreatment of glassy carbon electrode: the bare glassy carbon electrode of L-type, 3 mm of diameter, with 0.3 μm of Al of granularity2O3Suspension polishing, steams
Distilled water is rinsed well, then successively in HNO3, ethyl alcohol, be cleaned by ultrasonic in secondary distilled water, dry spare；
(2) Zn-Mg-Te/TiO for taking 6 μ L claims 2 to prepare2Clean glass carbon electricity obtained by composite material drop coating to step (1)
Pole surface is dried at room temperature, is cleaned with the PBS buffer solution of 0.01 pH=7.4 mol/L, is obtained Zn-Mg-Te/TiO2 /GCE
Modified electrode；
Take the squamous cell anticancrin anti-SCCA solution drop coating of 6 μ L, 10 μ g/mL to Zn-Mg-Te/TiO again2/ GCE modification electricity
Pole surface is dried at room temperature, is cleaned loose anti-SCCA with the PBS buffer solution of 0.01 pH=7.4 mol/L, is obtained
anti-SCCA/Zn-Mg-Te/TiO2/ GCE modified electrode；
Again by anti-SCCA/Zn-Mg-Te/TiO2/ GCE modified electrode is crosslinked 5 min in glutaraldehyde steam, with 0.01
The PBS buffer solution of pH=7.4 mol/L is cleaned, and is dried；0.5 h is impregnated, in 0.25wt% bovine serum albumin solution with envelope
Electrode surface nonspecific binding site that may be present is closed, with the PBS buffer solution of 0.01 pH=7.4 mol/L after taking-up
It cleans, dries, obtain squamous cell carcinoma photoelectricity immunosensor, be placed in 4 DEG C of refrigerators and save for use.
4. method preparation is compound based on water-soluble Zn-Mg-Te quantum dot/titanium dioxide nano-rod according to claim 3
Application of the squamous cell carcinoma photoelectricity immunosensor of material in qualitative and/or quantitative detection cervical carcinoma.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811572333.6A CN109709327A (en) | 2018-12-21 | 2018-12-21 | Photoelectricity immunosensor based on water-soluble Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811572333.6A CN109709327A (en) | 2018-12-21 | 2018-12-21 | Photoelectricity immunosensor based on water-soluble Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109709327A true CN109709327A (en) | 2019-05-03 |
Family
ID=66256064
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811572333.6A Withdrawn CN109709327A (en) | 2018-12-21 | 2018-12-21 | Photoelectricity immunosensor based on water-soluble Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109709327A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110470718A (en) * | 2019-09-19 | 2019-11-19 | 济南大学 | It is a kind of for detecting the preparation method of the optical electro-chemistry immunosensor of cardiac muscle troponin I |
| CN110702758A (en) * | 2019-09-27 | 2020-01-17 | 广西师范大学 | Method for enhancing luminous intensity of squamous cell carcinoma antigen in electrochemical luminescence detection |
| CN111073635A (en) * | 2019-12-12 | 2020-04-28 | 吉林大学 | Multilevel chiral luminescence reinforced composite material and preparation method thereof |
-
2018
- 2018-12-21 CN CN201811572333.6A patent/CN109709327A/en not_active Withdrawn
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110470718A (en) * | 2019-09-19 | 2019-11-19 | 济南大学 | It is a kind of for detecting the preparation method of the optical electro-chemistry immunosensor of cardiac muscle troponin I |
| CN110702758A (en) * | 2019-09-27 | 2020-01-17 | 广西师范大学 | Method for enhancing luminous intensity of squamous cell carcinoma antigen in electrochemical luminescence detection |
| CN111073635A (en) * | 2019-12-12 | 2020-04-28 | 吉林大学 | Multilevel chiral luminescence reinforced composite material and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN100520366C (en) | SERS biological probe and method for making same | |
| Nagl et al. | Improving surface and defect center chemistry of fluorescent nanodiamonds for imaging purposes—a review | |
| CN109709327A (en) | Photoelectricity immunosensor based on water-soluble Zn-Mg-Te quantum dot/titanium dioxide nano-rod composite material | |
| CN106645754B (en) | Photoelectricity immunosensor based on water-soluble Cd-Ag-Te quantum dots/nanogold composite material | |
| CN104133069B (en) | Preparation method for double-functional mark photo-electrochemical sensor and application | |
| CN102410994B (en) | Magnetic double-mode optical probe and preparation method thereof | |
| CN104569435B (en) | A kind of preparation method of unmarked Optical Electro-Chemistry alpha-fetoprotein immunosensor | |
| CN108287187A (en) | A kind of electrochemical luminescence sensor | |
| CN106290514B (en) | A kind of TiO based on silicon phthalocyanine functionalization2It is situated between and sees the aflatoxin optical electro-chemistry detection method of crystal | |
| CN109283235B (en) | Based on NSCQDs/Bi2S3Photoelectrochemical sensor and preparation method thereof | |
| CN106066324A (en) | A kind of preparation method and application of electroluminescent chemiluminescence biosensor label | |
| Wang et al. | A label-free electrochemiluminescence aptasensor for carcinoembryonic antigen detection based on electrodeposited ZnS–CdS on MoS 2 decorated electrode | |
| CN108828042A (en) | A kind of preparation method of the interlayer type optical electro-chemistry sensor of cTnI | |
| CN110346438B (en) | Based on PbS/Co3O4Preparation method of composite signal attenuation type photoelectrochemical immunosensor | |
| Han et al. | A photoelectrochemical immunosensor for detection of α-fetoprotein based on Au-ZnO flower-rod heterostructures | |
| CN107589166B (en) | One kind being based on TiO2It is situated between and sees the label-free type zearalenone optical electro-chemistry detection method and sensor of crystal | |
| CN101655473B (en) | Preparation method of nanogold immunoelectrode | |
| CN103837517B (en) | The preparation method of metallic film/nanometic zinc oxide rod array Fluorescence Increasing material | |
| CN106248656B (en) | One kind being based on Ni nanoparticle Co2O4The preparation method and application of the electrochemical luminescence immunosensor of@AD bifunctional catalyst | |
| CN104764737B (en) | A kind of monochromatic ECL immunologic detection methods based on green-light radiation quantum dot | |
| CN110133252A (en) | For detecting kit and detection method and its application of carcinomebryonic antigen | |
| Wang et al. | A visible light photoelectrochemical sensor for tumor marker detection using tin dioxide quantum dot–graphene as labels | |
| CN109142745B (en) | Preparation method and application of photoelectrochemical immunosensor based on stannic oxide/cadmium carbonate/cadmium sulfide | |
| CN103611946A (en) | Preparation method of gold nanocluster fluorescent materials protected by methionine | |
| CN109884029A (en) | Silver/graphite alkene quantum dot nano enzyme, SERS detection kit and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20190503 |