CN104880559A - Preparation method and application of Pt-carbon nitride/graphene carbon nitride tumor marker CA199 biological sensor - Google Patents
Preparation method and application of Pt-carbon nitride/graphene carbon nitride tumor marker CA199 biological sensor Download PDFInfo
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- CN104880559A CN104880559A CN201510248655.5A CN201510248655A CN104880559A CN 104880559 A CN104880559 A CN 104880559A CN 201510248655 A CN201510248655 A CN 201510248655A CN 104880559 A CN104880559 A CN 104880559A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
- G01N33/57488—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites involving compounds identifable in body fluids
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/70—Mechanisms involved in disease identification
- G01N2800/7023—(Hyper)proliferation
- G01N2800/7028—Cancer
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
- G01N33/57438—Specifically defined cancers of liver, pancreas or kidney
Abstract
The invention relates to a preparation method and application of a Pt-carbon nitride/graphene carbon nitride tumor marker CA199 biological sensor, and belongs to the technical field of novel functional materials and biological sensing detection. A Pt nano particle has a good catalytic performance to hydrogen peroxide, and the great specific surface area of carbon nitride/graphene is utilized for fixedly loading the Pt nano particle on the surface of the carbon nitride/graphene to act as a marker for detecting an antibody, so that the super-flexible detection of the pancreatic cancer tumor marker CA199 is realized, and has great significance to the early diagnosis and prognosis of the tumor marker.
Description
Technical field
The present invention relates to preparation method and the application of a kind of Pt-carbonitride/Graphene pancreatic tumour mark CA199 biology sensor.Specifically adopt Pt-carbonitride/Graphene as detection antibody labeling thing, prepare a kind of biology sensor detecting pancreatic tumour mark CA199, belong to new function material and bio-sensing detection technique field.
Background technology
Cancer of pancreas belongs to the higher tumour of the grade of malignancy of digestive system, and in recent years, the incidence of disease of cancer of pancreas increases to some extent, and thus, early diagnosis, early treatment is extremely important.Pancreatic tumour mark CA199 is mainly present in the tissue of stomach and intestine or Pancreas cancer patients, the Pancreas cancer patients in tumor in digestive tract, and in serum, CA199 content obviously raises, and the detection of CA199 is contributed to the early diagnosis of cancer of pancreas.
Nano material, because of the optics of its excellence, calorifics, electricity, mechanics and chemical property, makes it obtain in production, life and scientific research and applies widely.Graphene nano layer has large specific surface area, and catalytic performance is good, good biocompatibility, strengthens the advantages such as electron transmission.Pt is catalyst based has good catalytic performance to hydrogen peroxide, is widely used in battery, sensor field, and Pt nano particle and the material containing nitrogen element combine the activity that can improve material.The present invention using Pt-carbonitride/graphene composite material as detection antibody labeling thing, improve the sensitivity of biology sensor, selective good, the sensitive height of sensor that the present invention builds, detectability are low, simple operation and other advantages, overcome the deficiency of the methods such as radio immunoassay, enzyme-linked immunosorbent assay and kit.
Summary of the invention
An object of the present invention is the preparation of a kind of Pt-carbonitride/Graphene pancreatic tumour mark CA199 biology sensor.
Two of object of the present invention is by the detection of this biosensor application in pancreatic tumour mark CA199.
technical scheme of the present invention
The preparation of 1.Pt-carbonitride/Graphene
(1) preparation of graphene oxide
0.3 ~ 0.6g graphite and 1.8 ~ 3.6g potassium permanganate are added in the flask of 500 mL, by the H of 36 ~ 72 mL
2sO
4with the H of 4 ~ 8 mL
3pO
4add after mixing in flask, 50 DEG C add hot reflux 12 h, potpourri are poured on 40 ~ 80 mL on ice, add the hydrogen peroxide of 0.6 ~ 2 mL, stir 30 min, after centrifugal, use the hydrochloric acid of 0.2 mol/L, ethanol, washed with diethylether respectively, vacuum drying at 40 DEG C, obtained graphene oxide;
(2) preparation of polypyrrole/graphene oxide
The CTAB of 5 ~ 10 g is dissolved in the HNO of 80 mL, 1 mol/L
3in solution, under agitation, 0.05 ~ 0.2 g graphene oxide is added, at being heated to 100 DEG C, add 0.2 ~ 1 g pyrroles, continuous heating 60 min, add 1 ~ 3 g ammonium persulfate, react 180 min, potpourri after centrifugal, washing, vacuum drying at 40 DEG C, obtained polypyrrole/graphene oxide;
(3) preparation of antibody labeling thing-Pt-carbonitride/Graphene is detected
Under the atmosphere of argon gas, by polypyrrole/graphene oxide 800 DEG C calcining 2 h, obtained carbonitride/Graphene, by the K of 2 ~ 10 mg carbonitride/Graphenes and 1 ~ 3 mL, 3 mg/mL
2ptCl
4/ EG joins in 10 mL EG, and ultrasonic 0.5 h adds the NaOH/EG of 5 ~ 10 mL EG and 0.2 mL, 2 mol/L subsequently, under power is 300 W, microwave reaction 110 s, potpourri through centrifugal, washing after, vacuum drying at 60 DEG C, obtained Pt-carbonitride/Graphene.
2. detect antibody hatching thing-Pt-carbonitride/Graphene-Ab
2the preparation of solution
By the Pt-of 1 ~ 5 mg carbonitride/graphene dispersion in 1 mL ultrapure water, mix with the detection antibody-solutions of 1 mL, 5 ~ 12 μ g/mL, vibrate hatching 12 h at 4 DEG C, centrifuging, lower sediment be scattered in the pH=7.4 phosphate buffered solution of 1 mL, 1/15 mol/L, the obtained antibody that detects hatches thing-Pt-carbonitride/Graphene-Ab
2solution, saves backup at 4 DEG C.
3. the preparation method of Pt-carbonitride/Graphene pancreatic tumour mark CA199 biology sensor
(1) use the alundum (Al2O3) burnishing powder of 1.0,0.3,0.05 μm to glass-carbon electrode polishing successively, clean up with ultrapure water;
(2) get 6 μ L, the solution of 3-aminopropyl triethoxysilane functionalized graphene of 0.5 ~ 2 mg/mL is added drop-wise to electrode surface, dry under room temperature;
(3) by the pancreatic tumour mark CA199 capture antibody Ab of 6 μ L, 5 ~ 15 μ g/mL
1drop to electrode surface, dry at 4 DEG C, ultrapure water;
(4) drip 3 μ L, massfraction be the bovine serum albumin solution of 5 ~ 15 mg/mL to electrode surface, dry at 4 DEG C, ultrapure water;
(5) drip 6 μ L concentration be the pancreatic tumour mark CA199 solution of 0.0001 ~ 30 ng/mL to electrode surface, dry at 4 DEG C, ultrapure water;
(6) 4 ~ 6 μ L are detected antibody hatching thing-Pt-carbonitride/Graphene-Ab
2solution drops to electrode surface, is placed in 4 DEG C of refrigerators and hatches 1 h, after cleaning, dry, obtained a kind of Pt-carbonitride/Graphene pancreatic tumour mark CA199 biology sensor.
4. the detection method of pancreatic tumour mark CA199
(1) three-electrode system of electrochemical workstation is adopted to measure, using the immunosensor prepared by claim 1 as working electrode, saturated calomel electrode is to electrode, platinum electrode is auxiliary electrode, in the phosphate buffered solution of pH=7.4, during employing, the method for m-electric current scans, and input voltage is-0.4 V, working time 400 s;
(2) in the phosphate buffered solution of 10 mL, pH=7.4, the pancreatic tumour mark CA199 standard solution of 0.0001 ~ 30 ng/mL is tested, record current changes, linear according to the concentration of gained current differential and pancreatic tumour mark CA199, drawing curve;
(3) pancreatic tumour mark CA199 standard solution is replaced by testing sample solution to detect.
useful achievement of the present invention
(1) 3-aminopropyl triethoxysilane functionalized graphene has larger specific surface area, good electric conductivity and good biocompatibility and stability, in addition, 3-aminopropyl triethoxysilane functionalized graphene has a large amount of amino, its dispersiveness in water can not only be improved, and in conjunction with a large amount of capture antibodies, sensitivity and the stability of sensor can be added.
(2) Pt is catalyst based has good catalytic performance to hydrogen peroxide, is widely used in sensor field, and Pt nano particle and the material containing nitrogen element combine the activity that can improve material.
(3) electrochemical immunosensor prepared of the present invention is for the detection of pancreatic tumour mark, and the response time is short, and detectability is low, and the range of linearity is wide, can realize simple, quick, highly sensitive and specific detection.
Embodiment
embodiment 1the preparation of Pt-carbonitride/Graphene
(1) preparation of graphene oxide
0.3 g graphite and 1.8 g potassium permanganate are added in the flask of 500 mL, by the H of 36 mL
2sO
4with the H of 4 mL
3pO
4add in above-mentioned flask after mixing, 50 DEG C add hot reflux 12 h, potpourri are poured on 40 mL on ice, add the hydrogen peroxide of 0.6 mL, stir 30 min, after centrifugal, use the hydrochloric acid of 0.2 mol/L, ethanol, washed with diethylether respectively, vacuum drying at 40 DEG C, obtained graphene oxide;
(2) preparation of polypyrrole/graphene oxide
The CTAB of 5 g is dissolved in the HNO of 80 mL, 1 mol/L
3in solution, under agitation, 0.05 g graphene oxide is added, at being heated to 100 DEG C, add 0.2 g pyrroles, continuous heating 60 min, add 1 g ammonium persulfate, react 180 min, potpourri after centrifugal, washing, vacuum drying at 40 DEG C, obtained polypyrrole/graphene oxide;
(3) preparation of antibody labeling thing-Pt-carbonitride/Graphene is detected
Under the atmosphere of argon gas, by polypyrrole/graphene oxide 800 DEG C calcining 2 h, obtained carbonitride/Graphene, by the K of 2 mg carbonitride/Graphenes and 1 mL, 3 mg/mL
2ptCl
4/ EG joins in 10 mL EG, and ultrasonic 0.5 h adds the NaOH/EG of 5 mL EG and 0.2 mL, 2 mol/L subsequently, under power is 300 W, microwave reaction 110 s, potpourri through centrifugal, washing after, vacuum drying at 60 DEG C, obtained Pt-carbonitride/Graphene.
embodiment 2the preparation of Pt-carbonitride/Graphene
(1) preparation of graphene oxide
0.5 g graphite and 3.0 g potassium permanganate are added in the flask of 500 mL, by the H of 60 mL
2sO
4with the H of 6 mL
3pO
4add in above-mentioned flask after mixing, 50 DEG C add hot reflux 12 h, potpourri are poured on 60 mL on ice, add the hydrogen peroxide of 1 mL, stir 30 min, after centrifugal, use the hydrochloric acid of 0.2 mol/L, ethanol, washed with diethylether respectively, vacuum drying at 40 DEG C, obtained graphene oxide;
(2) preparation of polypyrrole/graphene oxide
The CTAB of 6 g is dissolved in the HNO of 80 mL, 1 mol/L
3in solution, under agitation, 0.1 g graphene oxide is added, at being heated to 100 DEG C, add 0.5 g pyrroles, continuous heating 60 min, add 2 g ammonium persulfates, react 180 min, potpourri after centrifugal, washing, vacuum drying at 40 DEG C, obtained polypyrrole/graphene oxide;
(3) preparation of antibody labeling thing-Pt-carbonitride/Graphene is detected
Under the atmosphere of argon gas, by polypyrrole/graphene oxide 800 DEG C calcining 2 h, obtained carbonitride/Graphene, by the K of 5 mg carbonitride/Graphenes and 2 mL, 3 mg/mL
2ptCl
4/ EG joins in 10 mL EG, and ultrasonic 0.5 h adds the NaOH/EG of 7 mL EG and 0.2 mL, 2 mol/L subsequently, under power is 300 W, microwave reaction 110 s, potpourri through centrifugal, washing after, vacuum drying at 60 DEG C, obtained Pt-carbonitride/Graphene.
embodiment 3the preparation of Pt-carbonitride/Graphene
(1) preparation of graphene oxide
0.6 g graphite and 3.6 g potassium permanganate are added in the flask of 500 mL, by the H of 72 mL
2sO
4with the H of 8 mL
3pO
4add in above-mentioned flask after mixing, 50 DEG C add hot reflux 12 h, potpourri are poured on 80 mL on ice, add the hydrogen peroxide of 2 mL, stir 30 min, after centrifugal, use the hydrochloric acid of 0.2 mol/L, ethanol, washed with diethylether respectively, vacuum drying at 40 DEG C, obtained graphene oxide;
(2) preparation of polypyrrole/graphene oxide
The CTAB of 10 g is dissolved in the HNO of 80 mL, 1 mol/L
3in solution, under agitation, 0.2 g graphene oxide is added, at being heated to 100 DEG C, add 1 g pyrroles, continuous heating 60 min, add 3 g ammonium persulfates, react 180 min, potpourri after centrifugal, washing, vacuum drying at 40 DEG C, obtained polypyrrole/graphene oxide;
(3) preparation of antibody labeling thing-Pt-carbonitride/Graphene is detected
Under the atmosphere of argon gas, by polypyrrole/graphene oxide 800 DEG C calcining 2 h, obtained carbonitride/Graphene, by the K of 10 mg carbonitride/Graphenes and 3 mL, 3 mg/mL
2ptCl
4/ EG joins in 10 mL EG, and ultrasonic 0.5 h adds the NaOH/EG of 10 mL EG and 0.2 mL, 2 mol/L subsequently, under power is 300 W, microwave reaction 110 s, potpourri through centrifugal, washing after, vacuum drying at 60 DEG C, obtained Pt-carbonitride/Graphene.
embodiment 4detect antibody hatching thing-Pt-carbonitride/Graphene-Ab
2the preparation of solution
By the Pt-of 1 mg carbonitride/graphene dispersion in 1 mL ultrapure water, mix with the detection antibody-solutions of 1 mL, 5 μ g/mL, vibrate hatching 12 h at 4 DEG C, centrifuging, lower sediment be scattered in the pH=7.4 phosphate buffered solution of 1 mL, 1/15 mol/L, the obtained antibody that detects hatches thing-Pt-carbonitride/Graphene-Ab
2solution, saves backup at 4 DEG C.
embodiment 5detect antibody hatching thing-Pt-carbonitride/Graphene-Ab
2the preparation of solution
By the Pt-of 2 mg carbonitride/graphene dispersion in 1 mL ultrapure water, mix with the detection antibody-solutions of 1 mL, 10 μ g/mL, vibrate hatching 12 h at 4 DEG C, centrifuging, lower sediment be scattered in the pH=7.4 phosphate buffered solution of 1 mL, 1/15 mol/L, the obtained antibody that detects hatches thing-Pt-carbonitride/Graphene-Ab
2solution, saves backup at 4 DEG C.
embodiment 6detect antibody hatching thing-Pt-carbonitride/Graphene-Ab
2the preparation of solution
By the Pt-of 5 mg carbonitride/graphene dispersion in 1 mL ultrapure water, mix with the detection antibody-solutions of 1 mL, 12 μ g/mL, vibrate hatching 12 h at 4 DEG C, centrifuging, lower sediment be scattered in the pH=7.4 phosphate buffered solution of 1 mL, 1/15 mol/L, the obtained antibody that detects hatches thing-Pt-carbonitride/Graphene-Ab
2solution, saves backup at 4 DEG C.
embodiment 7the preparation method of a kind of Pt-carbonitride/Graphene pancreatic tumour mark CA199 biology sensor
(1) use the alundum (Al2O3) burnishing powder of 1.0,0.3,0.05 μm to glass-carbon electrode polishing successively, clean up with ultrapure water;
(2) get 6 μ L, the solution of 3-aminopropyl triethoxysilane functionalized graphene of 0.5 mg/mL is added drop-wise to electrode surface, dry under room temperature;
(3) by the pancreatic tumour mark CA199 capture antibody Ab of 6 μ L, 5 μ g/mL
1drop to electrode surface, dry at 4 DEG C, ultrapure water;
(4) drip 3 μ L, massfraction be the bovine serum albumin solution of 5 mg/mL to electrode surface, dry at 4 DEG C, ultrapure water;
(5) drip 6 μ L concentration be the pancreatic tumour mark CA199 solution of 0.0001 ~ 30 ng/mL to electrode surface, dry at 4 DEG C, ultrapure water;
(6) 4 μ L are detected antibody hatching thing-Pt-carbonitride/Graphene-Ab
2solution drops to electrode surface, is placed in 4 DEG C of refrigerators and hatches 1 h, after cleaning, dry, obtained a kind of Pt-carbonitride/Graphene pancreatic tumour mark CA199 biology sensor.
embodiment 8the preparation method of a kind of Pt-carbonitride/Graphene pancreatic tumour mark CA199 biology sensor
(1) use the alundum (Al2O3) burnishing powder of 1.0,0.3,0.05 μm to glass-carbon electrode polishing successively, clean up with ultrapure water;
(2) get 6 μ L, the solution of 3-aminopropyl triethoxysilane functionalized graphene of 1 mg/mL is added drop-wise to electrode surface, dry under room temperature;
(3) by the pancreatic tumour mark CA199 capture antibody Ab of 6 μ L, 10 μ g/mL
1drop to electrode surface, dry at 4 DEG C, ultrapure water;
(4) drip 3 μ L, massfraction be the bovine serum albumin solution of 10 mg/mL to electrode surface, dry at 4 DEG C, ultrapure water;
(5) drip 6 μ L concentration be the pancreatic tumour mark CA199 solution of 0.0001 ~ 30 ng/mL to electrode surface, dry at 4 DEG C, ultrapure water;
(6) 5 μ L are detected antibody hatching thing-Pt-carbonitride/Graphene-Ab
2solution drops to electrode surface, is placed in 4 DEG C of refrigerators and hatches 1 h, after cleaning, dry, obtained a kind of Pt-carbonitride/Graphene pancreatic tumour mark CA199 biology sensor.
embodiment 9the preparation method of a kind of Pt-carbonitride/Graphene pancreatic tumour mark CA199 biology sensor
(1) use the alundum (Al2O3) burnishing powder of 1.0,0.3,0.05 μm to glass-carbon electrode polishing successively, clean up with ultrapure water;
(2) get 6 μ L, the solution of 3-aminopropyl triethoxysilane functionalized graphene of 2 mg/mL is added drop-wise to electrode surface, dry under room temperature;
(3) by the pancreatic tumour mark CA199 capture antibody Ab of 6 μ L, 15 μ g/mL
1drop to electrode surface, dry at 4 DEG C, ultrapure water;
(4) drip 3 μ L, massfraction be the bovine serum albumin solution of 15 mg/mL to electrode surface, dry at 4 DEG C, ultrapure water;
(5) drip 6 μ L concentration be the pancreatic tumour mark CA199 solution of 0.0001 ~ 30 ng/mL to electrode surface, dry at 4 DEG C, ultrapure water;
(6) 6 μ L are detected antibody hatching thing-Pt-carbonitride/Graphene-Ab
2solution drops to electrode surface, is placed in 4 DEG C of refrigerators and hatches 1 h, after cleaning, dry, obtained a kind of Pt-carbonitride/Graphene pancreatic tumour mark CA199 biology sensor.
embodiment 10the detection method of pancreatic tumour mark CA199
(1) three-electrode system of electrochemical workstation is adopted to measure, using the immunosensor prepared by claim 1 as working electrode, saturated calomel electrode is to electrode, platinum electrode is auxiliary electrode, in the phosphate buffered solution of pH=7.4, during employing, the method for m-electric current scans, and input voltage is-0.4 V, working time 400 s;
(2) in the phosphate buffered solution of 10 mL, pH=7.4, the pancreatic tumour mark CA199 standard solution of 0.0001 ~ 30 ng/mL is tested, record current changes, linear according to the concentration of gained current differential and pancreatic tumour mark CA199, drawing curve;
(3) replaced by testing sample solution pancreatic tumour mark CA199 standard solution to detect, recording the range of linearity is 0.0001 ~ 25 ng/mL, detects and is limited to 25 fg/mL.
Claims (4)
1. a preparation method for Pt-carbonitride/Graphene pancreatic tumour mark CA199 biology sensor, the preparation of Pt-carbonitride/Graphene, comprises the following steps:
(1) preparation of graphene oxide
0.3 ~ 0.6g graphite and 1.8 ~ 3.6g potassium permanganate are added in the flask of 500 mL, by the H of 36 ~ 72 mL
2sO
4with the H of 4 ~ 8 mL
3pO
4add after mixing in flask, 50 DEG C add hot reflux 12 h, potpourri are poured on 40 ~ 80 mL on ice, add the hydrogen peroxide of 0.6 ~ 2 mL, stir 30 min, after centrifugal, use the hydrochloric acid of 0.2 mol/L, ethanol, washed with diethylether respectively, vacuum drying at 40 DEG C, obtained graphene oxide;
(2) preparation of polypyrrole/graphene oxide
The CTAB of 5 ~ 10 g is dissolved in the HNO of 80 mL, 1 mol/L
3in solution, under agitation, 0.05 ~ 0.2 g graphene oxide is added, at being heated to 100 DEG C, add 0.2 ~ 1 g pyrroles, continuous heating 60 min, add 1 ~ 3 g ammonium persulfate, react 180 min, potpourri after centrifugal, washing, vacuum drying at 40 DEG C, obtained polypyrrole/graphene oxide;
(3) preparation of antibody labeling thing-Pt-carbonitride/Graphene is detected
Under the atmosphere of argon gas, by polypyrrole/graphene oxide 800 DEG C calcining 2 h, obtained carbonitride/Graphene, by the K of 2 ~ 10 mg carbonitride/Graphenes and 1 ~ 3 mL, 3 mg/mL
2ptCl
4/ EG joins in 10 mL EG, and ultrasonic 0.5 h adds the NaOH/EG of 5 ~ 10 mL EG and 0.2 mL, 2 mol/L subsequently, under power is 300 W, microwave reaction 110 s, potpourri through centrifugal, washing after, vacuum drying at 60 DEG C, obtained Pt-carbonitride/Graphene.
2. a preparation method for Pt-carbonitride/Graphene pancreatic tumour mark CA199 biology sensor, detects antibody hatching thing-Pt-carbonitride/Graphene-Ab
2the preparation of solution, comprises the following steps:
By the Pt-of 1 ~ 5 mg carbonitride/graphene dispersion in 1 mL ultrapure water, mix with the detection antibody-solutions of 1 mL, 5 ~ 12 μ g/mL, vibrate hatching 12 h at 4 DEG C, centrifuging, lower sediment be scattered in the pH=7.4 phosphate buffered solution of 1 mL, 1/15 mol/L, the obtained antibody that detects hatches thing-Pt-carbonitride/Graphene-Ab
2solution, saves backup at 4 DEG C.
3. a preparation method for Pt-carbonitride/Graphene pancreatic tumour mark CA199 biology sensor, is characterized in that, comprise the following steps:
(1) use the alundum (Al2O3) burnishing powder of 1.0,0.3,0.05 μm to glass-carbon electrode polishing successively, clean up with ultrapure water;
(2) get 6 μ L, the solution of 3-aminopropyl triethoxysilane functionalized graphene of 0.5 ~ 2 mg/mL is added drop-wise to electrode surface, dry under room temperature;
(3) by the pancreatic tumour mark CA199 capture antibody Ab of 6 μ L, 5 ~ 15 μ g/mL
1drop to electrode surface, dry at 4 DEG C, ultrapure water;
(4) drip 3 μ L, massfraction be the bovine serum albumin solution of 5 ~ 15 mg/mL to electrode surface, dry at 4 DEG C, ultrapure water;
(5) drip 6 μ L concentration be the pancreatic tumour mark CA199 solution of 0.0001 ~ 30 ng/mL to electrode surface, dry at 4 DEG C, ultrapure water;
(6) 4 ~ 6 μ L are detected antibody hatching thing-Pt-carbonitride/Graphene-Ab
2solution drops to electrode surface, is placed in 4 DEG C of refrigerators and hatches 1 h, after cleaning, dry, obtained a kind of Pt-carbonitride/Graphene pancreatic tumour mark CA199 biology sensor.
4. immunosensor prepared by the preparation method as described in claim 1 ~ 3, for the detection method of pancreatic tumour mark CA199, is characterized in that, comprises following analytical procedure:
(1) three-electrode system of electrochemical workstation is adopted to measure, using the immunosensor prepared by claim 1 as working electrode, saturated calomel electrode is to electrode, platinum electrode is auxiliary electrode, in the phosphate buffered solution of pH=7.4, during employing, the method for m-electric current scans, and input voltage is-0.4 V, working time 400 s;
(2) in the phosphate buffered solution of 10 mL, pH=7.4, the pancreatic tumour mark CA199 standard solution of 0.0001 ~ 30 ng/mL is tested, record current changes, linear according to the concentration of gained current differential and pancreatic tumour mark CA199, drawing curve;
(3) pancreatic tumour mark CA199 standard solution is replaced by testing sample solution to detect.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105301241A (en) * | 2015-10-22 | 2016-02-03 | 宁波大学 | Preparing method and application of sandwich type electrochemical luminescence immunosensor for detecting tumor marker |
| CN108690865A (en) * | 2018-05-25 | 2018-10-23 | 军事科学院军事医学研究院军事兽医研究所 | A kind of platinum-nano flower and its preparation method and application |
| WO2021213298A1 (en) * | 2020-04-20 | 2021-10-28 | 山东第一医科大学(山东省医学科学院) | Immunofluorescence kit for detecting ca199 expression of peripheral blood circulating tumor cells of pancreatic cancer patient and detection method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104297480A (en) * | 2014-09-26 | 2015-01-21 | 济南大学 | Preparation method and application of sandwich type immunosensor for prostate specific antigens |
| CN104459157A (en) * | 2014-12-17 | 2015-03-25 | 济南大学 | Preparation method and application of immunosensor based on ferrocene-cuprous oxide cubic nanometer frame mark |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104297480A (en) * | 2014-09-26 | 2015-01-21 | 济南大学 | Preparation method and application of sandwich type immunosensor for prostate specific antigens |
| CN104459157A (en) * | 2014-12-17 | 2015-03-25 | 济南大学 | Preparation method and application of immunosensor based on ferrocene-cuprous oxide cubic nanometer frame mark |
Non-Patent Citations (3)
| Title |
|---|
| CANCAN HUANG等: "Graphene based catalyst", 《ENERGY & ENVIRONMENTAL SCIENCE》, 20 August 2012 (2012-08-20) * |
| CHENGZHOU ZHU, SHAOJUN DONG: "Recent progress in grapheme-based nanomaterials as advanced electrocatalysts towards oxygen reduction reaction", 《NANOSCALE》, 3 January 2013 (2013-01-03) * |
| HAIBO WANG等: "Pd nanoparticles on carbon nitride-graphene for the selective electro-oxidation of glycerol in alkaline solution.", 《ACS CATALYSIS》, vol. 5, 17 April 2015 (2015-04-17) * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105301241A (en) * | 2015-10-22 | 2016-02-03 | 宁波大学 | Preparing method and application of sandwich type electrochemical luminescence immunosensor for detecting tumor marker |
| CN105301241B (en) * | 2015-10-22 | 2017-02-01 | 宁波大学 | Preparing method and application of sandwich type electrochemical luminescence immunosensor for detecting tumor marker |
| CN108690865A (en) * | 2018-05-25 | 2018-10-23 | 军事科学院军事医学研究院军事兽医研究所 | A kind of platinum-nano flower and its preparation method and application |
| WO2021213298A1 (en) * | 2020-04-20 | 2021-10-28 | 山东第一医科大学(山东省医学科学院) | Immunofluorescence kit for detecting ca199 expression of peripheral blood circulating tumor cells of pancreatic cancer patient and detection method |
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