CN108828042A - A kind of preparation method of the interlayer type optical electro-chemistry sensor of cTnI - Google Patents
A kind of preparation method of the interlayer type optical electro-chemistry sensor of cTnI Download PDFInfo
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Abstract
The present invention relates to a kind of preparation methods of the interlayer type optical electro-chemistry sensor of cTnI, belong to nano-functional material, immunoassay and optical electro-chemistry biosensor technique field.The positive square piece TiO of nanometer is sensitized with carboxylated CdS quantum dot2001 crystal face, enhance its visible absorption, obtain the composite material TiO that photoelectric activity significantly improves2CTnI antibody, bovine serum albumin(BSA) and cTnI antigen are assembled into TiO by LBL self-assembly method by/CdS2On/CdS composite material, Ag@Cu is utilized2O core-shell nano is secondary antibody marker, TiO2Specific binding between/CdS excellent photoelectric activity and cTnI antigen-antibody is realized to the super sensitivity detection of cTnI, has great importance for the analysis detection application of cTnI.
Description
Technical field
The invention belongs to nano-functional material, immunoassay and optical electro-chemistry biosensor technique fields, provide one
The preparation method of the interlayer type optical electro-chemistry sensor of kind cTnI.Specifically received using the modification of carboxylated CdS quantum dot
The positive square piece TiO of rice2001 crystal face as antibody capture substrate, use the Ag@Cu with double inhibition effect2O core-shell nano grain
Son is used as secondary antibody marker, prepares a kind of interlayer type optical electro-chemistry sensor for detecting cTnI specific antigen.
Background technique
Cardiovascular disease be the world today threaten human health number one killer, and acute myocardial infarction AMI have rapid onset,
The features such as disability rate is high, the death rate is high, therefore the research for acute myocardial infarction AMI is always hot spot.And cardiac troponin is international
The best marker of the sensitivity and specificity of the upper diagnosis acute myocardial infarction AMI generally acknowledged, popular feeling Troponin I is myocardium calcium egg
One of the subunit of white compound, Cardiac-specific and susceptibility with higher, referred to as clinical " gold standard ".Cause
This, the early diagnosis of cTnI is of great significance.
Cardiovascular disease be the world today threaten human health number one killer, and acute myocardial infarction AMI have rapid onset,
The features such as disability rate is high, the death rate is high, therefore the research for acute myocardial infarction AMI is always hot spot.And cardiac troponin is international
The best marker of the sensitivity and specificity of the upper diagnosis acute myocardial infarction AMI generally acknowledged, popular feeling Troponin I is myocardium calcium egg
One of the subunit of white compound, Cardiac-specific and susceptibility with higher, referred to as clinical " gold standard ".Cause
This, the early diagnosis of cTnI is of great significance.
At present, it has been reported that detection cTnI analysis method have enzyme-linked immunization, enzyme-linked fluorescence analysis,
Optomagnetic biosensor, enzyme mark electrophoresis and Electrical chemiluminescence immunoassay analysis method etc..Although there are many detection method of cTnI,
But these method and steps are complicated, take a long time, detection sensitivity is low, testing staff requires height.In order to overcome the above analysis side
The drawbacks of method, the present invention devise a kind of high specificity, and high sensitivity, selectivity is good, operates fast and convenient optical electro-chemistry and exempts from
Epidemic disease analysis method.
The present invention utilizes photoelectrochemical assay method, is sensitized TiO with carboxylated CdS quantum dot2001 crystal face, enhance it
Visible absorption obtains the composite material TiO that photoelectric activity significantly improves2/ CdS, by LBL self-assembly method, by myocardium calcium
Protein I antibody, bovine serum albumin(BSA) and cTnI antigen are assembled into TiO2On/CdS composite material, with Ag@Cu2O nucleocapsid
Nanoparticle is secondary antibody marker, utilizes Ag@Cu2Ag particle core is total to the plasma of visible light energy in O core-shell nano
Vibration transfer and p-type semiconductor Cu2O shell photo-generate electron-hole pairs compete electron donor ascorbic acid and the steric hindrance of itself
Increase the supported quantity of antibody, TiO with big specific surface area2/ CdS excellent photoelectric activity and cTnI antigen-antibody
Between specific binding, construct a kind of based on TiO2The interlayer type optical electro-chemistry sensor of the cTnI of/CdS.It should
Sensor has excellent optical electro-chemistry activity, and with high sensitivity, the range of linearity is wide, detection limit is low, detection is quick, prepared
The advantages that journey is relatively easy realizes the hypersensitive analysis to cTnI, mentions for effectively detection cTnI at present
New method is supplied.
Summary of the invention
The present invention provides a kind of preparation method of the interlayer type optical electro-chemistry sensor of cTnI, realize pair
The super sensitivity detection of cTnI.An object of the present invention is to provide a kind of interlayer type optical electro-chemistry of cTnI
The preparation method of sensor.The second object of the present invention is to prepared interlayer type optical electro-chemistry sensor be realized to cardiac muscle
The rapid sensitive of calcium protein I detects.
Technical scheme is as follows：
1. a kind of preparation method of the interlayer type optical electro-chemistry sensor of cTnI, steps are as follows：
（1）The ITO electro-conductive glass of the cm of 2.5 cm × 0.8 is successively used into dish washing liquid, acetone, dehydrated alcohol and ultrapure water ultrasound
Cleaning, is dried with nitrogen；
（2）Take the positive square piece titanium dioxide TiO of nanometer of 8 ~ 10 μ L, 8 ~ 12mg/mL2Hanging drop is added to conductive glass electrode
Conducting surface, naturally dry at room temperature；
（3）In TiO2The electrode surface of modification is added dropwise the cadmium sulfide CdS quantum dot solution of 4 ~ 5 μ L carboxylated, dries in the air at room temperature
Dry, TiO is made in ultrapure water electrode surface2/ CdS electrode；
（4）In TiO2The thioacetic acid of 4 ~ 5 μ L, 0.1 mol/L is added dropwise in/CdS electrode surface, dries at room temperature, continues to be added dropwise
1- ethyl -3- (3- dimethyl aminopropyl)-carbodiimides/n-hydroxysuccinimide of 4 ~ 5 μ L, reaction 20 ~ 40
Ultrapure water, naturally dry are used after min；
（5）Modified electrode surface continue 4 ~ 5 μ L of dropwise addition, concentration be 3 ~ 5 μ g/mL cTnI antibody-solutions,
Ultrapure water electrode surface is used after reacting 20 ~ 40 min, is dried in 4 DEG C of refrigerators；
（6）Continue 4 ~ 5 μ L of dropwise addition, the BSA solution that mass fraction is 1 ~ 1.5 %, with non-specific on enclosed-electrode surface
Active site is used ultrapure water electrode surface after reacting 20 ~ 40 min, is dried in 4 DEG C of refrigerators；
（7）Continue the cTnI antigenic solution of 3 ~ 5 μ L of dropwise addition, 0.00002 ~ 50 ng/mL, ultrapure water electricity
Pole surface is dried in 4 DEG C of refrigerators；
（8）Continue the Ag@Cu of 4 ~ 8 μ L of dropwise addition, 0.3 ~ 1.0 mg/mL2The cTnI of O core-shell nano label
The antibody-solutions of antigen are identified in electrode surface and antigentic specificity, are hatched 1 h at room temperature, are cleaned up, obtained one kind is based on
TiO2The interlayer type optical electro-chemistry sensor of the cTnI of/CdS, stores for future use in 4 DEG C of refrigerators.
1- ethyl -3- (the 3- dimethyl aminopropyl)-carbodiimides/n-hydroxysuccinimide contains 1 × 10-2 1- ethyl -3- (3- dimethyl aminopropyl)-carbodiimides and 2 × 10 of mol/L-3The n-hydroxysuccinimide of mol/L.
2. TiO2The preparation of/CdS, steps are as follows：
（1）0.6 ~ 1.0 mL hydrofluoric acid is added in 8 ~ 10 mL butyl titanates, stirs 15 ~ 30 min, acquired solution
It moves into polytetrafluoro liner autoclave, 180 DEG C of 22 ~ 26 h of reaction, is down to room temperature, centrifugation, washing, vacuum drying obtain
To TiO2；
（2）The thioacetic acid of 125 μ L is added drop-wise to the 0.01 mol/L CdCl of 25 mL2In aqueous solution, lead to nitrogen 20 ~ 40
Min is 11 with 1 mol/L sodium hydroxide tune pH value, the 0.1 mol/L sodium sulfide solution of 2.5 mL is slowly injected into solution
In, mixture reacts 4 h in the environment of nitrogen, obtains the CdS quantum dot of carboxylated, stores in 4 °C of refrigerators stand-by；
（3）Take the TiO of 8 ~ 10 μ L, 8 ~ 12 mg/mL2Hanging drop is added to the conducting surface of conductive glass electrode, at room temperature
Naturally dry continues the CdS quantum dot solution of 4 ~ 5 μ L carboxylated of dropwise addition, dries at room temperature, ultrapure water electrode surface,
TiO is made2/CdS。
3. Ag@Cu2The preparation of O core-shell nano, steps are as follows：
（1）The silver nitrate of 30 ~ 40mg is dissolved in the ultrapure water of 180 ~ 220 mL, and 30 min are stirred in 85 DEG C of reflux；
（2）?（1）The citric acid three sodium solution of 4 mL 1% is added in solution, boils 30 min, obtains silver solution；
（3）Under magnetic stirring, 10 ~ 12 g PVP are added to the 0.01 mol/L Cu (NO of 500 mL3)2In aqueous solution, to
After powder is completely dissolved, 40.0 mL silver solutions are added dropwise；
（4）340 μ L N are added immediately at room temperature2H4-H2O（35%）Solution, mixture stirs 5 min, clear with water and dehydrated alcohol
Wash Ag@Cu2O core-shell nano, then dispersed with dehydrated alcohol, it is stored for future use in 4 DEG C of refrigerators.
4. the detection method of cTnI specific antigen, steps are as follows：
（1）It is tested using the three-electrode system of electrochemical workstation, saturated calomel electrode is reference electrode, and platinum electrode is
Auxiliary electrode, prepared cTnI specific antigen interlayer type optical electro-chemistry sensor are working electrode, slow in PBS
It rushes in solution and is tested, PBS buffer solution is the ascorbic acid containing 0.1mol/L that 10 mL ~ 15 mL, pH are 5.0 ~ 8.5
Phosphate buffer solution；
（2）Using when m- current method cTnI specific antigen solution is detected, setting voltage is 0 V, operation
100 s of time, excitation light source are LED light, and working curve is drawn in record current variation；
（3）CTnI specific antigen solution is replaced to detect testing sample solution, according to step（2）With（3）In
Method detected, photo-current intensity according to responseIAnd working curve, obtain the content of sample to be tested center Troponin I.
The results show, m- current differential and cTnI specific antigen concentration are in 0.02 pg/ when of the present invention
Good linear relationship is kept within the scope of the ng/mL of mL ~ 50, detection limit reaches 6.7fg/mL.
Beneficial achievement of the invention
（1）The positive square piece titanium dioxide TiO of the nanometer that the present invention synthesizes2, wherein 001 crystal face accounts for 80% or more, there is good photoelectricity
Active, big surface area, the advantages that cheap and stability is good, can increase the load to functional material, improve material
Electric conductivity enhances the sensitivity of sensor.
（2）Carboxylated CdS quantum dot is sensitized TiO2001 crystal face, TiO can be enhanced2To the absorbability of visible light,
Obtain the TiO that photoelectric activity significantly improves2/ CdS composite material, the composite material can accelerate the separation of electron hole pair, increase
Add its electric conductivity, and further increases the photoelectric activity of sensor.
（3）Using Ag@Cu2O core-shell nano utilizes Ag@Cu as secondary antibody marker2Ag in O core-shell nano
Daughter nucleus is to the plasma resonance transfer of visible light energy and p-type semiconductor Cu2O shell photo-generate electron-hole pairs compete electron donor
（Ascorbic acid）And itself steric hindrance and big specific surface area increase the supported quantity of antibody, according to various concentration
Determinand combination secondary antibody marker inhibits the difference of photosignal intensity, improves the sensitivity of sensor, has widened linear model
It encloses, significantly reduces the detection limit of sensor, realize the super sensitivity detection to cTnI.
Specific embodiment
Now the present invention is further illustrated by specific embodiment, but not limited to this
A kind of preparation method of the interlayer type optical electro-chemistry sensor of the cTnI of embodiment 1
（1）The ITO electro-conductive glass of the cm of 2.5 cm × 0.8 is successively used into dish washing liquid, acetone, dehydrated alcohol and ultrapure water ultrasound
Cleaning, is dried with nitrogen；
（2）Take the positive square piece titanium dioxide TiO of nanometer of 8 μ L, 8 mg/mL2Hanging drop is added to the conduction of conductive glass electrode
Face, at room temperature naturally dry；
（3）In TiO2The electrode surface of modification is added dropwise the CdS quantum dot solution of 4 μ L carboxylated, dries at room temperature, ultrapure water punching
Electrode surface is washed, TiO is made2/ CdS electrode；
（4）In TiO2The thioacetic acid of 4 μ L, 0.1 mol/L is added dropwise in/CdS electrode surface, dries at room temperature, continues that 4 μ L are added dropwise
1- ethyl -3- (3- dimethyl aminopropyl)-carbodiimides/n-hydroxysuccinimide, react 30 min after use ultrapure water
It rinses, naturally dry；
（5）Continue the cTnI antibody-solutions for 4 μ L being added dropwise, concentration is 3 μ g/mL, reaction 30 on modified electrode surface
Ultrapure water electrode surface is used after min, is dried in 4 DEG C of refrigerators；
（6）Continue the BSA solution for 4 μ L being added dropwise, mass fraction is 1 %, with nonspecific activity site on enclosed-electrode surface,
Ultrapure water electrode surface is used after reacting 30 min, is dried in 4 DEG C of refrigerators；
（7）Continue the cTnI antigenic solution that 3 μ L, 0.00002 ~ 50 ng/mL is added dropwise, ultrapure water electrode table
Face is dried in 4 DEG C of refrigerators；
（8）Continue the Ag@Cu that 4 μ L, 0.4 mg/mL is added dropwise2The antibody of the cTnI antigen of O core-shell nano label
Solution is identified in electrode surface and antigentic specificity, is hatched 1 h at room temperature, is cleaned up, and is made a kind of and is based on TiO2/ CdS's
The interlayer type optical electro-chemistry sensor of cTnI, stores for future use in 4 DEG C of refrigerators.
1- ethyl -3- (the 3- dimethyl aminopropyl)-carbodiimides/n-hydroxysuccinimide contains 1 × 10-2
1- ethyl -3- (3- dimethyl aminopropyl)-carbodiimides and 2 × 10 of mol/L-3The N- hydroxysuccinimidyl acyl of mol/L is sub-
Amine.
A kind of preparation method of the interlayer type optical electro-chemistry sensor of the cTnI of embodiment 2
（1）The ITO electro-conductive glass of the cm of 2.5 cm × 0.8 is successively used into dish washing liquid, acetone, dehydrated alcohol and ultrapure water ultrasound
Cleaning, is dried with nitrogen；
（2）Take the positive square piece titanium dioxide TiO of nanometer of 9 μ L, 10 mg/mL2Hanging drop is added to the conduction of conductive glass electrode
Face, at room temperature naturally dry；
（3）In TiO2The electrode surface of modification is added dropwise the CdS quantum dot solution of 5 μ L carboxylated, dries at room temperature, ultrapure water punching
Electrode surface is washed, TiO is made2/ CdS electrode；
（4）In TiO2The thioacetic acid of 5 μ L, 0.1 mol/L is added dropwise in/CdS electrode surface, dries at room temperature, continues that 5 μ L are added dropwise
1- ethyl -3- (3- dimethyl aminopropyl)-carbodiimides/n-hydroxysuccinimide, react 30 min after use ultrapure water
It rinses, naturally dry；
（5）Continue the cTnI antibody-solutions for 5 μ L being added dropwise, concentration is 4 μ g/mL, reaction 40 on modified electrode surface
Ultrapure water electrode surface is used after min, is dried in 4 DEG C of refrigerators；
（6）Continue the BSA solution for 5 μ L being added dropwise, mass fraction is 1 %, with nonspecific activity site on enclosed-electrode surface,
Ultrapure water electrode surface is used after reacting 30 min, is dried in 4 DEG C of refrigerators；
（7）Continue the cTnI antigenic solution that 4 μ L, 0.00002 ~ 50 ng/mL is added dropwise, ultrapure water electrode table
Face is dried in 4 DEG C of refrigerators；
（8）The Ag@Cu of 6 μ L, 0.6 mg/mL is added dropwise2The antibody-solutions of the cTnI antigen of O core-shell nano label
It is identified in electrode surface and antigentic specificity, hatches 1 h at room temperature, clean up, be made a kind of and be based on TiO2The cardiac muscle of/CdS
The interlayer type optical electro-chemistry sensor of calcium protein I, stores for future use in 4 DEG C of refrigerators.
1- ethyl -3- (the 3- dimethyl aminopropyl)-carbodiimides/n-hydroxysuccinimide contains 1 × 10-2
1- ethyl -3- (3- dimethyl aminopropyl)-carbodiimides and 2 × 10 of mol/L-3The N- hydroxysuccinimidyl acyl of mol/L is sub-
Amine.
A kind of preparation method of the interlayer type optical electro-chemistry sensor of the cTnI of embodiment 3
（1）The ITO electro-conductive glass of the cm of 2.5 cm × 0.8 is successively used into dish washing liquid, acetone, dehydrated alcohol and ultrapure water ultrasound
Cleaning, is dried with nitrogen；
（2）Take the positive square piece titanium dioxide TiO of nanometer of 10 μ L, 12 mg/mL2Hanging drop is added to the conduction of conductive glass electrode
Face, at room temperature naturally dry；
（3）In TiO2The electrode surface of modification is added dropwise the CdS quantum dot solution of 5 μ L carboxylated, dries at room temperature, ultrapure water punching
Electrode surface is washed, TiO is made2/ CdS electrode；
（4）In TiO2The thioacetic acid of 5 μ L, 0.1 mol/L is added dropwise in/CdS electrode surface, dries at room temperature, continues that 5 μ L are added dropwise
1- ethyl -3- (3- dimethyl aminopropyl)-carbodiimides/n-hydroxysuccinimide, react 40 min after use ultrapure water
It rinses, naturally dry；
（5）Continue the cTnI antibody-solutions for 5 μ L being added dropwise, concentration is 5 μ g/mL, reaction 40 on modified electrode surface
Ultrapure water electrode surface is used after min, is dried in 4 DEG C of refrigerators；
（6）Continue the BSA solution for 4 μ L being added dropwise, mass fraction is 1.5 %, with nonspecific activity position on enclosed-electrode surface
Point is used ultrapure water electrode surface after reacting 40 min, is dried in 4 DEG C of refrigerators；
（7）Continue the cTnI antigenic solution that 5 μ L, 0.00002 ~ 50 ng/mL is added dropwise, ultrapure water electrode table
Face is dried in 4 DEG C of refrigerators；
（8）Continue the Ag@Cu that 5 μ L, 0.8 mg/mL is added dropwise2The antibody of the cTnI antigen of O core-shell nano label
Solution is identified in electrode surface and antigentic specificity, is hatched 1 h at room temperature, is cleaned up, and is made a kind of and is based on TiO2/ CdS's
The interlayer type optical electro-chemistry sensor of cTnI, stores for future use in 4 DEG C of refrigerators.
1- ethyl -3- (the 3- dimethyl aminopropyl)-carbodiimides/n-hydroxysuccinimide contains 1 × 10-2
1- ethyl -3- (3- dimethyl aminopropyl)-carbodiimides and 2 × 10 of mol/L-3The N- hydroxysuccinimidyl acyl of mol/L is sub-
Amine.
4 TiO of embodiment2The preparation of/CdS, steps are as follows：
（1）0.6 mL hydrofluoric acid is added in 8 mL butyl titanates, stirs 15 min, acquired solution moves into polytetrafluoro liner
In autoclave, 180 DEG C of 24 h of reaction are down to room temperature, and centrifugation, washing, vacuum drying obtain TiO2；
（2）The thioacetic acid of 125 μ L is added drop-wise to the 0.01 mol/L CdCl of 25 mL2In aqueous solution, lead to 20 min of nitrogen,
It is 11 with 1 mol/L sodium hydroxide tune pH value, the 0.1 mol/L sodium sulfide solution of 2.5 mL is slowly injected into solution, mixes
It closes object and reacts 4 h in the environment of nitrogen, obtain the CdS quantum dot of carboxylated, stored in 4 °C of refrigerators stand-by；
（3）Take the TiO of 10 μ L, 8 mg/mL2Hanging drop is added to the conducting surface of conductive glass electrode, at room temperature naturally dry,
Continue the CdS quantum dot solution that 4 μ L carboxylated are added dropwise, dry at room temperature, ultrapure water electrode surface, TiO is made2 /
CdS。
5 TiO of embodiment2The preparation of/CdS, steps are as follows：
（1）0.8 mL hydrofluoric acid is added in 9 mL butyl titanates, stirs 20 min, acquired solution moves into polytetrafluoro liner
In autoclave, 26 h of heating reaction at 180 DEG C are down to room temperature, and centrifugation, washing, vacuum drying obtain TiO2；
（2）The thioacetic acid of 125 μ L is added drop-wise to the 0.01 mol/L CdCl of 25 mL2In aqueous solution, lead to 30 min of nitrogen,
It is 11 with 1 mol/L sodium hydroxide tune pH value, the 0.1 mol/L sodium sulfide solution of 2.5 mL is slowly injected into solution, mixes
It closes object and reacts 4 h in the environment of nitrogen, obtain the CdS quantum dot of carboxylated, stored in 4 °C of refrigerators stand-by；
（3）Take the TiO of 10 μ L, 10 mg/mL2Hanging drop is added to the conducting surface of conductive glass electrode, at room temperature naturally dry,
Continue the CdS quantum dot solution that 5 μ L carboxylated are added dropwise, dry at room temperature, ultrapure water electrode surface, TiO is made2/CdS。
6 TiO of embodiment2The preparation of/CdS, steps are as follows：
（1）1.0 mL hydrofluoric acid are added in 10 mL butyl titanates, stir 30 min, acquired solution moves into polytetrafluoro liner
In autoclave, 24 h of heating reaction at 180 DEG C are down to room temperature, and centrifugation, washing, vacuum drying obtain TiO2；
（2）The thioacetic acid of 125 μ L is added drop-wise to the 0.01 mol/L CdCl of 25 mL2In aqueous solution, lead to 40 min of nitrogen,
It is 11 with 1 mol/L sodium hydroxide tune pH value, the 0.1 mol/L sodium sulfide solution of 2.5 mL is slowly injected into solution, mixes
It closes object and reacts 4 h in the environment of nitrogen, obtain the CdS quantum dot of carboxylated, stored in 4 °C of refrigerators stand-by；
（3）Take the TiO of 10 μ L, 12 mg/mL2Hanging drop is added to the conducting surface of conductive glass electrode, at room temperature naturally dry,
Continue the CdS quantum dot solution that 4 μ L carboxylated are added dropwise, dry at room temperature, ultrapure water electrode surface, TiO is made2/CdS。
7 Ag@Cu of embodiment2The preparation of O core-shell nano, steps are as follows：
（1）The silver nitrate of 30 mg is dissolved in the ultrapure water of 180 mL, and 30 min are stirred in 85 DEG C of reflux；
（2）?（1）The citric acid three sodium solution of 4 mL 1% is added in solution, boils 30 min, obtains silver solution；
（3）Under magnetic stirring, 10 g PVP are added to the 0.01 mol/L Cu (NO of 500 mL3)2In aqueous solution, to powder
After being completely dissolved, 40.0 mL silver solutions are added dropwise；
（4）340 μ L N are added immediately at room temperature2H4-H2O（35%）Solution, mixture stirs 5 min, clear with water and dehydrated alcohol
Wash Ag@Cu2O core-shell nano, then dispersed with dehydrated alcohol, it is stored for future use in 4 DEG C of refrigerators.
8 Ag@Cu of embodiment2The preparation of O core-shell nano, steps are as follows：
（1）The silver nitrate of 36 mg is dissolved in the ultrapure water of 200 mL, and 30 min are stirred in 85 DEG C of reflux；
（2）?（1）The citric acid three sodium solution of 4 mL 1% is added in solution, boils 30 min, obtains silver solution；
（3）Under magnetic stirring, 10.5 g PVP are added to the 0.01 mol/L Cu (NO of 500 mL3)2In aqueous solution, to powder
After end is completely dissolved, 40.0 mL silver solutions are added dropwise；
（4）340 μ L N are added immediately at room temperature2H4-H2O（35%）Solution, mixture stirs 5 min, clear with water and dehydrated alcohol
Wash Ag@Cu2O core-shell nano, then dispersed with dehydrated alcohol, it is stored for future use in 4 DEG C of refrigerators.
9 Ag@Cu of embodiment2The preparation of O core-shell nano, steps are as follows：
（1）The silver nitrate of 40 mg is dissolved in the ultrapure water of 220 mL, and 30 min are stirred in 85 DEG C of reflux；
（2）?（1）The citric acid three sodium solution of 4 mL 1% is added in solution, boils 30 min, obtains silver solution；
（3）Under magnetic stirring, 11 g PVP are added to the 0.01 mol/L Cu (NO of 500 mL3)2In aqueous solution, to powder
After being completely dissolved, 40.0 mL silver solutions are added dropwise；
（4）340 μ L N are added immediately at room temperature2H4-H2O（35%）Solution, mixture stirs 5 min, clear with water and dehydrated alcohol
Wash Ag@Cu2O core-shell nano, then dispersed with dehydrated alcohol, it is stored for future use in 4 DEG C of refrigerators.
The detection of 10 cTnI specific antigen of embodiment
（1）It is tested using the three-electrode system of electrochemical workstation, saturated calomel electrode is reference electrode, and platinum electrode is
Auxiliary electrode, prepared cTnI specific antigen interlayer type optical electro-chemistry sensor are working electrode, slow in PBS
It rushes in solution and is tested；
（2）Using when m- current method cTnI specific antigen solution is detected, setting voltage is 0 V, operation
100 s of time, excitation light source are LED light, and working curve is drawn in record current variation；
（3）CTnI specific antigen solution is replaced to detect testing sample solution, according to step（2）With（3）In
Method detected, photo-current intensity according to responseIAnd working curve, obtain the content of sample to be tested center Troponin I.
The PBS buffer solution is that 10 mL ~ 15 mL, pH are 5.0 ~ 8.5 containing 0.1 mol/L ascorbic acid
Phosphate buffer solution.
Claims (4)
1. a kind of preparation method of the interlayer type optical electro-chemistry sensor of cTnI, which is characterized in that steps are as follows：
（1）The ITO electro-conductive glass of the cm of 2.5 cm × 0.8 is successively used into dish washing liquid, acetone, dehydrated alcohol and ultrapure water ultrasound
Cleaning, is dried with nitrogen；
（2）Take the positive square piece titanium dioxide TiO of nanometer of 8 ~ 10 μ L, 8 ~ 12 mg/mL2Hanging drop is added to electro-conductive glass electricity
The conducting surface of pole, at room temperature naturally dry；
（3）In TiO2The electrode surface of modification is added dropwise the cadmium sulfide CdS quantum dot solution of 4 ~ 5 μ L carboxylated, dries in the air at room temperature
Dry, TiO is made in ultrapure water electrode surface2/ CdS electrode；
（4）In TiO2The thioacetic acid of 4 ~ 5 μ L, 0.1 mol/L is added dropwise in/CdS electrode surface, dries at room temperature, continues dropwise addition 4
1- ethyl -3- (3- dimethyl aminopropyl)-carbodiimides/n-hydroxysuccinimide of ~ 5 μ L reacts 20 ~ 40 min
Ultrapure water, naturally dry are used afterwards；
（5）Modified electrode surface continue 4 ~ 5 μ L of dropwise addition, concentration be 3 ~ 5 μ g/mL cTnI antibody-solutions,
Ultrapure water electrode surface is used after reacting 20 ~ 40 min, is dried in 4 DEG C of refrigerators；
（6）Continue 4 ~ 5 μ L of dropwise addition, the BSA solution that mass fraction is 1 ~ 1.5 %, with non-specific on enclosed-electrode surface
Active site is used ultrapure water electrode surface after reacting 20 ~ 40 min, is dried in 4 DEG C of refrigerators；
（7）Continue the cTnI antigenic solution of 3 ~ 5 μ L of dropwise addition, 0.00002 ~ 50 ng/mL, ultrapure water electricity
Pole surface is dried in 4 DEG C of refrigerators；
（8）Continue the Ag@Cu of 4 ~ 8 μ L of dropwise addition, 0.3 ~ 1.0 mg/mL2The cTnI of O core-shell nano label
The antibody-solutions of antigen are identified in electrode surface and antigentic specificity, are hatched 1 h at room temperature, are cleaned up, obtained one kind is based on
TiO2The interlayer type optical electro-chemistry sensor of the cTnI of/CdS, stores for future use in 4 DEG C of refrigerators；
1- ethyl -3- (the 3- dimethyl aminopropyl)-carbodiimides/n-hydroxysuccinimide contains 1 × 10-2
1- ethyl -3- (3- dimethyl aminopropyl)-carbodiimides and 2 × 10 of mol/L-3The N- hydroxysuccinimidyl acyl of mol/L is sub-
Amine.
2. a kind of preparation method of the interlayer type optical electro-chemistry sensor of cTnI as described in claim 1, described
TiO2The preparation of/CdS, which is characterized in that steps are as follows：
（1）0.6 ~ 1.0 mL hydrofluoric acid is added in 8 ~ 10 mL butyl titanates, stirs 15 ~ 30 min, acquired solution is moved
Enter in polytetrafluoro liner autoclave, 180 DEG C of 22 ~ 26 h of reaction, be down to room temperature, centrifugation, washing, vacuum drying obtain
TiO2；
（2）The thioacetic acid of 125 μ L is added drop-wise to the 0.01 mol/L CdCl of 25 mL2In aqueous solution, lead to nitrogen 20 ~ 40
Min is 11 with 1 mol/L sodium hydroxide tune pH value, the 0.1 mol/L sodium sulfide solution of 2.5 mL is slowly injected into solution
In, mixture reacts 4 h in the environment of nitrogen, obtains the CdS quantum dot of carboxylated, stores in 4 °C of refrigerators stand-by；
（3）Take the TiO of 8 ~ 10 μ L, 8 ~ 12mg/mL2Hanging drop is added to the conducting surface of conductive glass electrode, at room temperature certainly
It so dries, continues the CdS quantum dot solution of 4 ~ 5 μ L carboxylated of dropwise addition, dry at room temperature, ultrapure water electrode surface, make
Obtain TiO2/CdS。
3. a kind of preparation method of the interlayer type optical electro-chemistry sensor of cTnI as described in claim 1, described
Ag@Cu2The preparation of O core-shell nano, which is characterized in that steps are as follows：
（1）The silver nitrate of 30 ~ 40 mg is dissolved in the ultrapure water of 180 ~ 220 mL, and 30 min are stirred in 85 DEG C of reflux；
（2）?（1）The citric acid three sodium solution of 4 mL 1% is added in solution, boils 30 min, obtains silver solution；
（3）Under magnetic stirring, 10 ~ 12 g PVP are added to the 0.01 mol/L Cu (NO of 500 mL3)2In aqueous solution, to
After powder is completely dissolved, 40.0 mL silver solutions are added dropwise；
（4）340 μ L N are added immediately at room temperature2H4-H2O（35%）Solution, mixture stirs 5 min, clear with water and dehydrated alcohol
Wash Ag@Cu2O core-shell nano, then dispersed with dehydrated alcohol, it is stored for future use in 4 DEG C of refrigerators.
4. a kind of preparation method of the interlayer type optical electro-chemistry sensor of cTnI as described in claim 1, feature
It is, for the detection of cTnI specific antigen, detecting step is as follows：
（1）It is tested using the three-electrode system of electrochemical workstation, saturated calomel electrode is reference electrode, and platinum electrode is
Auxiliary electrode, prepared cTnI specific antigen interlayer type optical electro-chemistry sensor are working electrode, slow in PBS
It rushes in solution and is tested, PBS buffer solution is that 10 mL ~ 15 mL, pH are 5.0 ~ 8.5 containing 0.1 mol/L Vitamin C
The phosphate buffer solution of acid；
（2）Using when m- current method cTnI specific antigen solution is detected, setting voltage is 0 V, operation
100 s of time, excitation light source are LED light, and working curve is drawn in record current variation；
（3）CTnI specific antigen solution is replaced to detect testing sample solution, according to step（2）With（3）In
Method detected, photo-current intensity according to responseIAnd working curve, obtain the content of sample to be tested center Troponin I.
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