CN106248750B - A kind of fibrin ferment aptamer optical electro-chemistry sensor based on poly-dopamine complex capsule mark - Google Patents
A kind of fibrin ferment aptamer optical electro-chemistry sensor based on poly-dopamine complex capsule mark Download PDFInfo
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- G01N27/3278—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction involving nanosized elements, e.g. nanogaps or nanoparticles
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Abstract
The present invention relates to a kind of preparation method and application of the fibrin ferment aptamer optical electro-chemistry sensor based on poly-dopamine complex capsule mark, belong to preparation and the biology sensor detection technique field of new function nano composite material.High capacity characteristic based on poly-dopamine capsule, Cd is adsorbed with poly-dopamine capsule2+, obtain Cd2+@poly-dopamine complex capsules, for marking fibrin ferment aptamer；Using growth in situ method on poly-dopamine complex capsule in-situ preparation CdS nano-particles；Utilize TiO2The specific recognition function of cooperative effect and aptamer and object between the CdS in poly-dopamine complex capsule, success construction cost is cheap, interlayer type optical electro-chemistry immunosensor easy to operate, realizes the highly sensitive detection of fibrin ferment.
Description
Technical field
The present invention relates to a kind of preparation of the fibrin ferment aptamer sensor based on poly-dopamine complex capsule mark
Method and application, belong to preparation and the biology sensor detection technique field of new function nano composite material.
Background technology
The fibrin ferment formed by serine proteinases hydrolysis is a kind of important serine protease.In Human Physiology and disease
All played an important role during reason, the biological marker that can be diagnosed as relevant disease.Weigh the important of clotting mechanism
Index is the concentration and activity of fibrin ferment, for disclose tumour mechanism and as early diagnosis, treatment, curative effect and more after
Basis for estimation it is significant.
Optical electro-chemistry immunosensor is a kind of more novelty, the sensor quickly grown, and it has had photochemical method concurrently
With the advantage of electrochemical techniques；Meanwhile aptamer is come out by SELEX technology screenings, the adaptation that will be screened
Body is combined as recognition component with optical electro-chemistry sensor, has the high selectivity of aptamers and the advantage of specific binding,
Again have optical electro-chemistry sensor small volume, selectivity and sensitivity it is high, simple to operate, inexpensive the advantages that.Present invention system
For a kind of fibrin ferment aptamer optical electro-chemistry sensor based on poly-dopamine complex capsule mark.
The present invention prepares TiO using solvent-thermal method2Nano material, Cd is adsorbed using poly-dopamine capsule2+Obtain Cd2+@gathers
Dopamine complex capsule, for marking fibrin ferment aptamer, by growth in situ mode on poly-dopamine capsule it is in situ
CdS nano-particles are generated, due to TiO2Cooperative effect between generated in-situ CdS shows very excellent optical electro-chemistry
Activity, the sensitivity of sensor is improved, has widened the range of linearity, significantly reduce the detection limit of sensor, realized pair
The super sensitivity detection of fibrin ferment.This method combination optical electro-chemistry and aptamer technology, have high sensitivity, specificity it is good,
The advantages that simple operation, effectively overcome the deficiency of current blood coagulation enzyme assay method.
The content of the invention
An object of the present invention is to utilize poly-dopamine capsule absorption Cd2+, obtain Cd2+@poly-dopamine complex capsules,
Using growth in situ method, the in-situ preparation CdS nano-particles on poly-dopamine capsule.
The second object of the present invention is to be based on TiO2Cooperative effect and aptamer between CdS on poly-dopamine capsule
With the specific recognition function of object, a kind of interlayer type optical electro-chemistry immunosensor is constructed, realizes the oversoul of fibrin ferment
Quick detection.
Technical scheme is as follows：
A kind of 1. preparation of the fibrin ferment aptamer optical electro-chemistry sensor based on poly-dopamine complex capsule mark
（1）1 ~ 3 mL 1 mol/L nitric acid cadmium solution, fully vibration, centrifugation are added dropwise in poly-dopamine capsule solution
Supernatant liquor is removed, obtains Cd2+@poly-dopamine complex capsules；2 mL fibrin ferment aptamer solution are added, in 4 °C of constant temperature
Vibrate and 2 ~ 6 h are vibrated in case, centrifugation removes supernatant, obtains Cd2+The blood coagulation enzymatic nucleic acid adaptation of@poly-dopamines complex capsule mark
Body；The bovine serum albumin(BSA) BSA solution of 2 mL mass fraction 1% is added, 1 h is vibrated under normal temperature, is centrifuged off supernatant；Again
Ultra-pure water 2 mL is added, it is fully dispersed, obtain Cd2+The fibrin ferment aptamer solution of@poly-dopamines complex capsule mark, 4 °
C refrigerator storages are standby；
（2）By 2.5 × 0.8 cm2ITO electrode be immersed in 50 °C of acetone, water, second alcohol and water ultrasound respectively successively
30 min are cleaned, are then dried up with high pure nitrogen；10 μ L 1 ~ 6 mg/mL TiO is added dropwise2Nanoparticle suspension is in ITO electricity
Pole, after 300 ~ 600 °C of calcinings of Muffle furnace, obtain TiO2The ITO electrode of Nanoparticle Modified, is cooled to room temperature；
（3）In TiO2In the ITO electrode of Nanoparticle Modified, 2 ~ 7 μm of ol/L blood coagulation enzymatic nucleic acids that 6 μ L are added dropwise successively are fitted
Part, the bovine serum albumin(BSA) of mass fraction 1%, nonspecific binding site is closed, ultrapure water, is dried in 4 °C of refrigerators；
（4）It is added dropwise 1 × 10-15~1×10-11A series of thrombin solution of various concentrations of mol/L, 30 min are reacted, surpassed
Pure water rinsing, dry in 4 °C of refrigerators；
（5）6 μ L Cd is added dropwise2+The fibrin ferment aptamer solution of@poly-dopamines complex capsule mark is to electrode table
Face, 20 ~ 40 min are reacted, ultrapure water, are dried in 4 °C of refrigerators；
（6）6 μ L 0.1 ~ 1.0 mol/L Na is added dropwise2S solution, 10 ~ 30 min are reacted, with ultrapure water, 4 °C of ice
Dried in case, a kind of fibrin ferment aptamer optical electro-chemistry sensing based on poly-dopamine complex capsule mark has been prepared
Device.
2. the preparation of poly-dopamine capsule
（1）By the polystyrene microsphere alcohol dispersion liquid that 1 ~ 5 mL quality volume fraction is 2.5% and a diameter of 300 nm
5 ~ 15 min of emulsification in OP-10 emulsifying agents are added to, are subsequently dispersed the 80 mL 10 mmol/L trihydroxy methyl ammonia of pH 8.5
In methylmethane-hydrochloric acid buffer solution, the min of ultrasonic disperse 10；20 ~ 60 mg dopamines are added, at room temperature magnetic agitation reaction 2
~ 48 h, centrifuge, three times, ethanol is washed three times, obtains poly-dopamine polystyrene microsphere for washing, is disperseed standby in ethanol
With；
（2）The poly-dopamine@polystyrene microspheres prepared dissolving is distributed in 50 ~ 200 mL tetrahydrofurans, magnetic force
The h of stirring reaction 6 ~ 48, centrifuge, then distinguish centrifuge washing three times with tetrahydrofuran, ethanol and water successively, centrifugation terminates
After remove supernatant liquor, obtain poly-dopamine capsule, add ultra-pure water to disperse.
3. TiO2The preparation of nano-particle
0.01 ~ 0.03 mol tetra-butyl orthotitanates are taken to be dissolved in 20 ~ 40 mL ethanol solutions, then under magnetic stirring
10 mL ultra-pure water is added dropwise dropwise into above-mentioned solution, after 30 °C are reacted 60 ~ 180 min, suspension is transferred to polytetrafluoroethyl-ne
In the autoclave of alkene liner, and 2 ~ 24 h are reacted under the conditions of being maintained at 100 ~ 200 °C；Prepared sediment is collected, and respectively
It is each three times with absolute ethyl alcohol and the thorough centrifuge washing of ultra-pure water, 24 h are then dried in 50 °C of vacuum drying chambers, are pulverized
End is standby.
4. the detection method of fibrin ferment
（1）Detected using three-electrode system, the optical electro-chemistry sensor as prepared by claim 1 is work electricity
Pole, saturated calomel electrode are reference electrode, and platinum electrode is auxiliary electrode, is detected using optical electro-chemistry work station, photoelectricity
Detection uses i-t means of testing, and bias is arranged to 0 V, and light source is white light, and switch lamp is carried out every 10 s；
（2）In pHs 7.4 of 10 mL containing 0.1 mol/L ascorbic acid PBS cushioning liquid, worked by optical electro-chemistry
Stand and detect 1 × 10-15~1×10-11A series of thrombin standard solution of various concentrations of mol/L, by before and after recording switch lamp
Caused different photo-signals, drawing curve；
（3）Testing sample solution is detected instead of thrombin standard solution, the result of detection can pass through working curve
Check in.
The useful achievement of the present invention
（1）The high-load performance of poly-dopamine, is advantageous to Cd2+Load, obtained Cd2+@poly-dopamine complex capsules,
So as to be prepared for the marker material of novelty.
（2）Based on TiO2Cooperative effect on base material and poly-dopamine complex capsule between the CdS of growth in situ, drop
It is low to reduce test limit, realize super sensitivity detection.
A kind of fibrin ferment aptamer optical electro-chemistry sensor based on poly-dopamine complex capsule mark of embodiment 1
Prepare
（1）1 mL 1 mol/L nitric acid cadmium solution is added dropwise in poly-dopamine capsule solution, fully vibration, centrifugation removes
Supernatant liquor is removed, obtains Cd2+@poly-dopamine complex capsules；2 mL fibrin ferment aptamer solution are added, are shaken in 4 °C of constant temperature
Swing and 2 h are vibrated in case, centrifugation removes supernatant, obtains Cd2+The fibrin ferment aptamer of@poly-dopamines complex capsule mark；
The bovine serum albumin(BSA) BSA solution of 2 mL mass fraction 1% is added, 1 h is vibrated under normal temperature, is centrifuged off supernatant；Add
Ultra-pure water 2 mL, it is fully dispersed, obtain Cd2+The fibrin ferment aptamer solution of@poly-dopamines complex capsule mark, 4 °C of ice
Case stores for future use；
（2）By 2.5 × 0.8 cm2ITO electrode be immersed in 50 °C of acetone, water, second alcohol and water ultrasound respectively successively
30 min are cleaned, are then dried up with high pure nitrogen；10 μ L 1 mg/mL TiO is added dropwise2Nanoparticle suspension is in ITO electricity
Pole, after 300 °C of calcinings of Muffle furnace, obtain TiO2The ITO electrode of Nanoparticle Modified, is cooled to room temperature；
（3）In TiO2In the ITO electrode of Nanoparticle Modified, 6 μ L 2 μm of ol/L blood coagulations enzymatic nucleic acids adaptation is added dropwise successively
Body, the bovine serum albumin(BSA) of mass fraction 1%, nonspecific binding site is closed, ultrapure water, is dried in 4 °C of refrigerators；
（4）It is added dropwise 1 × 10-15~1×10-11A series of thrombin solution of various concentrations of mol/L, 30 min are reacted, surpassed
Pure water rinsing, dry in 4 °C of refrigerators；
（5）6 μ L Cd is added dropwise2+The fibrin ferment aptamer solution of@poly-dopamines complex capsule mark is to electrode table
Face, 20 min are reacted, ultrapure water, are dried in 4 °C of refrigerators；
（6）6 μ L 0.1 mol/L Na is added dropwise2S solution, 10 min are reacted, with ultrapure water, dried in the air in 4 °C of refrigerators
It is dry, a kind of fibrin ferment aptamer optical electro-chemistry sensor based on poly-dopamine complex capsule mark has been prepared.
A kind of fibrin ferment aptamer optical electro-chemistry sensor based on poly-dopamine complex capsule mark of embodiment 2
Preparation
（1）2 mL 1 mol/L nitric acid cadmium solution is added dropwise in poly-dopamine capsule solution, fully vibration, centrifugation removes
Supernatant liquor is removed, obtains Cd2+@poly-dopamine complex capsules；2 mL fibrin ferment aptamer solution are added, are shaken in 4 °C of constant temperature
Swing and 4 h are vibrated in case, centrifugation removes supernatant, obtains Cd2+The fibrin ferment aptamer of@poly-dopamines complex capsule mark；
The bovine serum albumin(BSA) BSA solution of 2 mL mass fraction 1% is added, 1 h is vibrated under normal temperature, is centrifuged off supernatant；Add
Ultra-pure water 2 mL, it is fully dispersed, obtain Cd2+The fibrin ferment aptamer solution of@poly-dopamines complex capsule mark, 4 °C of ice
Case stores for future use；
（2）By 2.5 × 0.8 cm2ITO electrode be immersed in 50 °C of acetone, water, second alcohol and water ultrasound respectively successively
30 min are cleaned, are then dried up with high pure nitrogen；10 μ L 4 mg/mL TiO is added dropwise2Nanoparticle suspension is in ITO electricity
Pole, after 450 °C of calcinings of Muffle furnace, obtain TiO2The ITO electrode of Nanoparticle Modified, is cooled to room temperature；
（3）In TiO2In the ITO electrode of Nanoparticle Modified, 6 μ L 5 μm of ol/L blood coagulations enzymatic nucleic acids adaptation is added dropwise successively
Body, the bovine serum albumin(BSA) of mass fraction 1%, nonspecific binding site is closed, ultrapure water, is dried in 4 °C of refrigerators；
（4）It is added dropwise 1 × 10-15~1×10-11A series of thrombin solution of various concentrations of mol/L, 30 min are reacted, surpassed
Pure water rinsing, dry in 4 °C of refrigerators；
（5）6 μ L Cd is added dropwise2+The fibrin ferment aptamer solution of@poly-dopamines complex capsule mark is to electrode table
Face, 30 min are reacted, ultrapure water, are dried in 4 °C of refrigerators；
（6）6 μ L 0.05 mol/L Na is added dropwise2S solution, 20 min are reacted, with ultrapure water, dried in the air in 4 °C of refrigerators
It is dry, a kind of fibrin ferment aptamer optical electro-chemistry sensor based on poly-dopamine complex capsule mark has been prepared.
A kind of fibrin ferment aptamer optical electro-chemistry sensor based on poly-dopamine complex capsule mark of embodiment 3
Prepare
（1）3 mL 1 mol/L nitric acid cadmium solution is added dropwise in poly-dopamine capsule solution, fully vibration, centrifugation removes
Supernatant liquor is removed, obtains Cd2+@poly-dopamine complex capsules；2 mL fibrin ferment aptamer solution are added, are shaken in 4 °C of constant temperature
Swing and 6 h are vibrated in case, centrifugation removes supernatant, obtains Cd2+The fibrin ferment aptamer of@poly-dopamines complex capsule mark；
The bovine serum albumin(BSA) BSA solution of 2 mL mass fraction 1% is added, 1 h is vibrated under normal temperature, is centrifuged off supernatant；Add
Ultra-pure water 2 mL, it is fully dispersed, obtain Cd2+The fibrin ferment aptamer solution of@poly-dopamines complex capsule mark, 4 °C of ice
Case stores for future use；
（2）By 2.5 × 0.8 cm2ITO electrode be immersed in 50 °C of acetone, water, second alcohol and water ultrasound respectively successively
30 min are cleaned, are then dried up with high pure nitrogen；10 μ L 6 mg/mL TiO is added dropwise2Nanoparticle suspension is in ITO electricity
Pole, after 600 °C of calcinings of Muffle furnace, obtain TiO2The ITO electrode of Nanoparticle Modified, is cooled to room temperature；
（3）In TiO2In the ITO electrode of Nanoparticle Modified, 6 μ L 7 μm of ol/L blood coagulations enzymatic nucleic acids adaptation is added dropwise successively
Body, the bovine serum albumin(BSA) of mass fraction 1%, nonspecific binding site is closed, ultrapure water, is dried in 4 °C of refrigerators；
（4）It is added dropwise 1 × 10-15~1×10-11A series of thrombin solution of various concentrations of mol/L, 30 min are reacted, surpassed
Pure water rinsing, dry in 4 °C of refrigerators；
（5）6 μ L Cd is added dropwise2+The fibrin ferment aptamer solution of@poly-dopamines complex capsule mark is to electrode table
Face, 40 min are reacted, ultrapure water, are dried in 4 °C of refrigerators；
（6）6 μ L 1.0 mol/L Na is added dropwise2S solution, 30 min are reacted, with ultrapure water, dried in the air in 4 °C of refrigerators
It is dry, a kind of fibrin ferment aptamer optical electro-chemistry sensor based on poly-dopamine complex capsule mark has been prepared.
The preparation of the poly-dopamine capsule of embodiment 4
（1）The polystyrene microsphere alcohol dispersion liquid that 1 mL quality volume fraction is 2.5% and a diameter of 300 nm is added
Enter into OP-10 emulsifying agents and emulsify 5 min, be subsequently dispersed the 80 mL 10 mmol/L trihydroxy methyl amino first of pH 8.5
In alkane-hydrochloric acid buffer solution, the min of ultrasonic disperse 10；20 mg dopamines are added, magnetic agitation reacts 2 h, centrifugation at room temperature
Separation, three times, ethanol is washed three times, obtains poly-dopamine polystyrene microsphere for washing, is disperseed standby in ethanol；
（2）The poly-dopamine@polystyrene microspheres prepared dissolving is distributed in 50 mL tetrahydrofurans, magnetic agitation
6 h are reacted, are centrifuged, then distinguish centrifuge washing three times with tetrahydrofuran, ethanol and water successively, centrifugation removes after terminating
Layer clear liquid, obtains poly-dopamine capsule, adds ultra-pure water to disperse.
The preparation of the poly-dopamine capsule of embodiment 5
（1）By the polystyrene microsphere alcohol dispersion liquid that 2.5 mL quality volume fraction is 2.5% and a diameter of 300 nm
It is added in OP-10 emulsifying agents and emulsifies 10 min, is subsequently dispersed the 80 mL 10 mmol/L trihydroxy methyl amino of pH 8.5
In methane-hydrochloric acid buffer solution, the min of ultrasonic disperse 10；40 mg dopamines are added, magnetic agitation reacts 24 h at room temperature,
Centrifuge, three times, ethanol is washed three times, obtains poly-dopamine polystyrene microsphere for washing, is disperseed standby in ethanol；
（2）The poly-dopamine@polystyrene microspheres prepared dissolving is distributed in 100 mL tetrahydrofurans, magnetic agitation
24 h are reacted, are centrifuged, then distinguish centrifuge washing three times with tetrahydrofuran, ethanol and water successively, centrifugation removes after terminating
Supernatant liquor, poly-dopamine capsule is obtained, adds ultra-pure water to disperse.
The preparation of the poly-dopamine capsule of embodiment 6
（1）The polystyrene microsphere alcohol dispersion liquid that 5 mL quality volume fraction is 2.5% and a diameter of 300 nm is added
Enter into OP-10 emulsifying agents and emulsify 15 min, be subsequently dispersed the 80 mL 10 mmol/L trihydroxy methyl amino first of pH 8.5
In alkane-hydrochloric acid buffer solution, the min of ultrasonic disperse 10；60 mg dopamines are added, magnetic agitation reacts 48 h at room temperature, from
The heart is separated, and three times, ethanol is washed three times, obtains poly-dopamine polystyrene microsphere for washing, is disperseed standby in ethanol；
（2）The poly-dopamine@polystyrene microspheres prepared dissolving is distributed in 200 mL tetrahydrofurans, magnetic agitation
48 h are reacted, are centrifuged, then distinguish centrifuge washing three times with tetrahydrofuran, ethanol and water successively, centrifugation removes after terminating
Supernatant liquor, poly-dopamine capsule is obtained, adds ultra-pure water to disperse.
Embodiment 7
TiO2The preparation of nano-particle
0.01 mol tetra-butyl orthotitanates are taken to be dissolved in 20 mL ethanol solutions, then under magnetic stirring to above-mentioned molten
10 mL ultra-pure water is added dropwise in liquid dropwise, after 30 °C are reacted 60 min, suspension is transferred to the high pressure of polytetrafluoroethyllining lining
In kettle, and 2 h are reacted under the conditions of being maintained at 100 °C；Prepared sediment is collected, and it is thorough with absolute ethyl alcohol and ultra-pure water respectively
Bottom centrifuge washing is each three times, and 24 h are then dried in 50 °C of vacuum drying chambers, and grind into powder is standby.
Embodiment 8
TiO2The preparation of nano-particle
0.02 mol tetra-butyl orthotitanates are taken to be dissolved in 30 mL ethanol solutions, then under magnetic stirring to above-mentioned molten
10 mL ultra-pure water is added dropwise in liquid dropwise, after 30 °C are reacted 120 min, suspension is transferred to the height of polytetrafluoroethyllining lining
Press in kettle, and 12 h are reacted under the conditions of being maintained at 150 °C；Collect prepared sediment, and respectively with absolute ethyl alcohol and ultrapure
The thorough centrifuge washing of water is each three times, and 24 h are then dried in 50 °C of vacuum drying chambers, and grind into powder is standby.
Embodiment 9
TiO2The preparation of nano-particle
0.03 mol tetra-butyl orthotitanates are taken to be dissolved in 40 mL ethanol solutions, then under magnetic stirring to above-mentioned molten
10 mL ultra-pure water is added dropwise in liquid dropwise, after 30 °C are reacted 180 min, suspension is transferred to the height of polytetrafluoroethyllining lining
Press in kettle, and 24 h are reacted under the conditions of being maintained at 200 °C；Collect prepared sediment, and respectively with absolute ethyl alcohol and ultrapure
The thorough centrifuge washing of water is each three times, and 24 h are then dried in 50 °C of vacuum drying chambers, and grind into powder is standby.
Embodiment 10
The detection method of fibrin ferment
（1）Detected using three-electrode system, the optical electro-chemistry sensor as prepared by claim 1 is work electricity
Pole, saturated calomel electrode are reference electrode, and platinum electrode is auxiliary electrode, is detected using optical electro-chemistry work station, photoelectricity
Detection uses i-t means of testing, and bias is arranged to 0 V, and light source is white light, and switch lamp is carried out every 10 s；
（2）In pHs 7.4 of 10 mL containing 0.1 mol/L ascorbic acid PBS cushioning liquid, worked by optical electro-chemistry
Stand and detect 1 × 10-15~1×10-11A series of thrombin standard solution of various concentrations of mol/L, by before and after recording switch lamp
Caused different photo-signals, drawing curve；
（3）Testing sample solution is detected instead of thrombin standard solution, the result of detection can pass through working curve
Check in.
Claims (3)
1. a kind of fibrin ferment aptamer optical electro-chemistry sensor based on poly-dopamine complex capsule mark, its feature exist
In comprising the following steps：
（1）1 ~ 3 mL 1 mol/L nitric acid cadmium solution, fully vibration are added dropwise in poly-dopamine capsule solution, is centrifuged off
Supernatant liquor, obtain Cd2+@poly-dopamine complex capsules；2 mL fibrin ferment aptamer solution are added, in 4 °C of constant temperature oscillations
2 ~ 6 h are vibrated in case, centrifugation removes supernatant, obtains Cd2+The fibrin ferment aptamer of@poly-dopamines complex capsule mark；
The bovine serum albumin(BSA) BSA solution of 2 mL mass fraction 1% is added, 1 h is vibrated under normal temperature, is centrifuged off supernatant；Add
Ultra-pure water 2 mL, it is fully dispersed, obtain Cd2+The fibrin ferment aptamer solution of@poly-dopamines complex capsule mark, 4 °C of ice
Case stores for future use；
（2）By 2.5 × 0.8 cm2ITO electrode be immersed in successively in 50 °C of acetone, water, second alcohol and water and be cleaned by ultrasonic 30 respectively
Min, then dried up with high pure nitrogen；10 μ L 1 ~ 6 mg/mL TiO is added dropwise2Nanoparticle suspension uses horse in ITO electrode
Not after 300 ~ 600 °C of calcinings of stove, TiO is obtained2The ITO electrode of Nanoparticle Modified, is cooled to room temperature；
（3）In TiO2In the ITO electrode of Nanoparticle Modified, 6 μ L 2 ~ 7 μm of ol/L fibrin ferment aptamers are added dropwise successively,
The bovine serum albumin(BSA) of mass fraction 1%, nonspecific binding site is closed, ultrapure water, is dried in 4 °C of refrigerators；
（4）It is added dropwise 1 × 10-15~1×10-11A series of thrombin solution of various concentrations of mol/L, react 30 min, ultra-pure water
Rinse, dried in 4 °C of refrigerators；
（5）6 μ L Cd is added dropwise2+The fibrin ferment aptamer solution of@poly-dopamines complex capsule mark is to electrode surface, instead
Answer 20 ~ 40 min, ultrapure water, dry in 4 °C of refrigerators；
（6）6 μ L 0.1 ~ 1.0 mol/L Na is added dropwise2S solution, 10 ~ 30 min are reacted, with ultrapure water, in 4 °C of refrigerators
Dry, a kind of fibrin ferment aptamer optical electro-chemistry sensor based on poly-dopamine complex capsule mark has been prepared；
The preparation method of described poly-dopamine capsule comprises the following steps：
（1）The polystyrene microsphere alcohol dispersion liquid that 1 ~ 5 mL quality volume fraction is 2.5% and a diameter of 300 nm is added
5 ~ 15 min are emulsified into OP-10 emulsifying agents, are subsequently dispersed the 80 mL 10 mmol/L trihydroxy methyl amino first of pH 8.5
In alkane-hydrochloric acid buffer solution, the min of ultrasonic disperse 10；20 ~ 60 mg dopamines are added, at room temperature magnetic agitation reaction 2 ~ 48
H, centrifuge, three times, ethanol is washed three times, obtains poly-dopamine polystyrene microsphere for washing, is disperseed standby in ethanol；
（2）The poly-dopamine@polystyrene microspheres prepared dissolving is distributed in 50 ~ 200 mL tetrahydrofurans, magnetic agitation
6 ~ 48 h are reacted, are centrifuged, then distinguish centrifuge washing three times with tetrahydrofuran, ethanol and water successively, centrifugation removes after terminating
Supernatant liquor is removed, poly-dopamine capsule is obtained, adds ultra-pure water to disperse.
A kind of 2. fibrin ferment aptamer optical electro-chemistry based on poly-dopamine complex capsule mark as claimed in claim 1
Sensor, described TiO2Nano-particle, it is characterised in that comprise the following steps：
0.01 ~ 0.03 mol tetra-butyl orthotitanates are taken to be dissolved in 20 ~ 40 mL ethanol solutions, it is then upward under magnetic stirring
The ultra-pure water that 10 mL are added dropwise in solution dropwise is stated, after 30 °C are reacted 60 ~ 180 min, suspension is transferred in polytetrafluoroethylene (PTFE)
In the autoclave of lining, and 2 ~ 24 h are reacted under the conditions of being maintained at 100 ~ 200 °C；Prepared sediment is collected, and uses nothing respectively
Water-ethanol and the thorough centrifuge washing of ultra-pure water are each three times, 24 h are then dried in 50 °C of vacuum drying chambers, grind into powder is standby
With.
A kind of 3. fibrin ferment aptamer optical electro-chemistry based on poly-dopamine complex capsule mark as claimed in claim 1
Sensor is used for the detection method of fibrin ferment, it is characterised in that comprises the following steps：
（1）Detected using three-electrode system, the optical electro-chemistry sensor as prepared by claim 1 is working electrode, is satisfied
It is reference electrode with calomel electrode, platinum electrode is auxiliary electrode, is detected using optical electro-chemistry work station, Photoelectric Detection is adopted
With i-t means of testing, bias is arranged to 0 V, and light source is white light, and switch lamp is carried out every 10 s；
（2）In pHs 7.4 of 10 mL containing 0.1 mol/L ascorbic acid PBS cushioning liquid, examined by optical electro-chemistry work station
Survey 1 × 10-15~1×10-11A series of thrombin standard solution of various concentrations of mol/L, produced by recording before and after switch lamp
Different photo-signals, drawing curve；
（3）Testing sample solution is detected instead of thrombin standard solution, the result of detection can looking into by working curve
.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610841305.4A CN106248750B (en) | 2016-09-22 | 2016-09-22 | A kind of fibrin ferment aptamer optical electro-chemistry sensor based on poly-dopamine complex capsule mark |
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| CN106645350B (en) * | 2016-12-29 | 2019-03-08 | 西南大学 | Poly-dopamine modifies application of the n-type semiconductor in building photoelectricity immunosensor |
| CN108918872A (en) * | 2018-07-25 | 2018-11-30 | 济南大学 | A kind of construction method for the photic electrochemical immunosensor of paper base detecting fibrin ferment |
| CN108918873B (en) * | 2018-07-30 | 2021-06-04 | 济南大学 | Preparation method and application of photoelectrochemical thrombin aptamer sensor based on dual inhibition of PS @ Au on ZnCdS |
| CN109975272B (en) * | 2019-03-13 | 2021-11-16 | 中国科学院苏州生物医学工程技术研究所 | Dopamine detection sensor and detection method based on electrochemiluminescence energy transfer |
| CN110488021B (en) * | 2019-08-21 | 2023-06-13 | 浙江理工大学 | Silk fibroin electrochemical immunosensor based on polydopamine-titanium dioxide composite material modified glassy carbon electrode |
| CN110596219B (en) * | 2019-10-17 | 2021-12-17 | 南京工业大学 | Photoelectrochemical gene sensor for detecting lung cancer marker and preparation method and application thereof |
| CN110865074B (en) * | 2019-11-21 | 2022-05-17 | 陕西科技大学 | Photonic crystal heavy metal sensor with immobilized enzyme and preparation method thereof |
| CN117309847B (en) * | 2023-11-28 | 2024-01-26 | 成都理工大学 | Trinitrotoluene detection sensor based on magnetic core-shell nanospheres |
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