CN104833806A - Preparation method and application of malignant tumour specific growth factor (TSGF) antigen electrogenerated chemiluminescence sensor - Google Patents
Preparation method and application of malignant tumour specific growth factor (TSGF) antigen electrogenerated chemiluminescence sensor Download PDFInfo
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- CN104833806A CN104833806A CN201510241947.6A CN201510241947A CN104833806A CN 104833806 A CN104833806 A CN 104833806A CN 201510241947 A CN201510241947 A CN 201510241947A CN 104833806 A CN104833806 A CN 104833806A
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Abstract
The invention relates to a preparation method and application of a malignant tumour specific growth factor (TSGF) antigen electrogenerated chemiluminescence sensor. The invention in particular relates to preparation and application of an electrogenerated chemiluminescence immunosensor taking aminated graphene GS-NH2 and an incubation substance of a detection antibody Ab2 as detection antibody labels and provides a simple, rapid and pollution-free preparation method and an application of a nitrogen doped carbon quantum dot electrogenerated chemiluminescence immunosensor. Linear range of the nitrogen doped carbon quantum dot electrogenerated chemiluminescence immunosensor is 0.01ng/mL-100ng/mL, and detection limit is 3.3pg/mL, so that rapid and sensitive detection on malignant TSGF antigen is realized.
Description
Technical field
The present invention relates to a kind of preparation method and application of Tumor specific factor TSGF antigen Electrochemiluminescsensor sensor.Be specifically related to a kind of nitrogen-doped carbon quantum dot as luminescent material, amination Graphene GS-NH
2with detection antibody A b
2hatching thing as the preparation and application of electrochemiluminescimmunosensor immunosensor detecting antibody labeling thing, belong to electrochemiluminescence detection technique field.
Background technology
TSGF has broad spectrum activity as the diagnosis of novel tumor markers to lung cancer, susceptibility, specificity are higher, and it has certain values in the auxiliary diagnosis of lung cancer.Tumor specific factor (TSGF) be malignant tumour formed and growth time impel tumour and periphery capillary breed in a large number and are discharged into the factor in peripheral blood, it is not only with the specificity of malignant tumour, and to be just discharged in blood at the initial period that cancerous swelling is formed and to acquire a certain degree.It has wide spectrum, susceptibility and specificity high to have bibliographical information.
According to the literature, the sensitivity of CEA to lung cancer only has 61.5%, β-M sensitivity also to only have 72.1%.The sensitivity of this testing result Sera of Lung Cancer TSGF is 81.2%, apparently higher than above-mentioned tumor marker.The detection of visible TSGF has broad spectrum activity, susceptibility, specific advantage, has very important meaning to the early diagnosis of lung cancer.
The early detection of lung cancer, early diagnosis, early treatment is one of important means improving its cure rate.Current topmost diagnostic method has physics method, histocytology method and chemical method three major types.Front two class methods are applicable to middle and advanced stage patient, and complicated operation, somewhat expensive; Chemical method is able to widespread use owing to having simple and easy to do advantage, and the fast development of particularly tumor marker detection in recent years, this method is one of Main Means becoming diagnosing early malignant tumor.
The present invention devises a kind of high specificity, highly sensitive, no radioactivity pollute, operates fast and convenient electrogenerated chemiluminescence immune analysis method.Use nitrogen-doped carbon quantum dot N-CQDs as base material in the present invention, amination Graphene with detect antibody and formed and hatch thing and build sandwich type immunosensor.The concentration of Tumor specific factor TSGF antigen is different, and the electroluminescence intensity that chemiluminescence detector detects is different, thus realizes the quantitative detection to Tumor specific factor TSGF antigen.The chemiluminescence immunoassay sensor of sending a telegraph built in the present invention has simple to operate, and cost is low, no radioactivity pollute, highly sensitive, the advantages such as high specificity, overcomes the some shortcomings of radio immunoassay and enzyme-linked immunosorbent assay.
Summary of the invention
The object of the invention is the problem of the detection method for existing Tumor specific factor TSGF antigen, a kind of preparation method and application of simple, quick, free of contamination nitrogen-doped carbon quantum dot electrochemiluminescimmunosensor immunosensor are provided, realize detecting the rapid sensitive of Tumor specific factor TSGF antigen
technical scheme of the present invention, comprises the following steps
1. the preparation method of a Tumor specific factor TSGF antigen Electrochemiluminescsensor sensor
(1) by diameter be the glass-carbon electrode Al of 4mm
2o
3burnishing powder is polished, and ultrapure water cleans up;
(2) the nitrogen-doped carbon quantum dot N-CQDs solution of 6 μ L, 8 ~ 12 μ g/mL is dripped to electrode surface, ultrapure water, dry under room temperature;
(3) when electrode is closely dry, drip 3 μ L, 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimides EDC that concentration is 0.3 ~ 0.5 mol/L and 0.05 ~ 0.2 mol/L N-hydroxy-succinamide NHS solution is to electrode surface, hatch 30min in 4 DEG C of refrigerators, dry under room temperature;
(4) drip 6 μ L, massfraction be the bovine serum albumin(BSA) BSA solution of 0.5 ~ 2.0% in electrode surface, with enclosed-electrode nonspecific activity site on the surface, ultrapure water, dries in 4 DEG C of refrigerators; Drip the Tumor specific factor TSGF antigen standard solution of a series of variable concentrations of 6 μ L, 10ng/mL ~ 100 ng/mL in electrode surface, ultrapure water, dry in 4 DEG C of refrigerators;
(5) the detection antibody hatching thing GS-NH of painting 6 μ L, 1 ~ 3 mg/mL is dripped
2@Ab
2solution, in electrode surface, is placed in 4 DEG C of refrigerators and dries, obtained a kind of Tumor specific factor TSGF antigen Electrochemiluminescsensor sensor.
2. prepared by nitrogen-doped carbon quantum dot N-CQDs
The diethyl pentetic acid DTPA of 0.2 ~ 0.4 g is dissolved in the ultrapure water of 10 ~ 30mL, ultrasonic disperse, dissolves at continuing to be heated to 230 DEG C, continues heated solvent evaporation, there is white solid, show the generation of nitrogen-doped carbon quantum dot N-CQDs when white solid becomes brown color; Be dissolved in 20mL ultrapure water by the thick product of gained, 8000 rpmin are centrifugal, and the freeze drying of gained supernatant liquor obtains pure nitrogen-doped carbon quantum dot N-CQDs solid, is ground into powder, obtained nitrogen-doped carbon quantum dot N-CQDs.
3. detect antibody hatching thing GS-NH
2@Ab
2prepared by solution
By the amination Graphene GS-NH of 1mL, 1 ~ 3 mg/mL
2solution, the detection antibody A b of 1mL, 5 ~ 10 μ g/mL
2solution joins in the centrifuge tube of 5 mL, and concussion 12h, makes amination Graphene GS-NH
2with detection antibody A b
2bonding, centrifuging, being distributed to 1mL, pH is in the phosphate buffer solution of 6.5 ~ 8.5, and the obtained antibody that detects hatches thing GS-NH
2@Ab
2solution.
The detection method of Tumor specific factor TSGF antigen
(1) three-electrode system of electrochemical workstation is used to test, Ag/AgCl electrode is as contrast electrode, platinum electrode is to electrode, prepared Electrochemiluminescsensor sensor is working electrode, electrochemical workstation and chemiluminescence detector are linked together the high pressure of photomultiplier is set to 650 V, scanning voltage is set to-2 ~ 0 V, sweep speed 0.1 V/s;
(2) 10 mL, pH 7.5 containing 80 mmol/L potassium persulfates PBS buffer solution in, by the Tumor specific factor TSGF antigen standard solution of electrogenerated chemiluminescence systems axiol-ogy to a series of variable concentrations of 0.01 ng/mL ~ 100 ng/mL, the electrochemiluminescence signal intensity produced, drawing curve;
(3) Tumor specific factor TSGF antigen standard solution is replaced by testing sample solution to detect.
useful achievement of the present invention
(1) electrochemiluminescimmunosensor immunosensor preparation method, with nitrogen-doped carbon quantum dot for luminescent material, utilizes the optical property that it is good, and the sensor of structure has higher sensitivity;
(2) detection antibody hatching thing GS-NH is utilized
2@Ab
2to the quenching effect of nitrogen-doped carbon quantum dot light emitting, build immunosensor;
(3) electrochemiluminescimmunosensor immunosensor prepared of the present invention is for the detection of Tumor specific factor TSGF antigen, and simple to operate, no radioactivity pollute, signal response wide ranges, can realize simple, quick, highly sensitive detection.Recording the range of linearity is 0.01 ng/mL ~ 100 ng/mL, detects and is limited to 3.3 pg/mL.
Specific embodiments
embodiment 1a kind of preparation method of Tumor specific factor TSGF antigen Electrochemiluminescsensor sensor:
1. the preparation method of a Tumor specific factor TSGF antigen Electrochemiluminescsensor sensor
(1) by diameter be the glass-carbon electrode Al of 4mm
2o
3burnishing powder is polished, and ultrapure water cleans up;
(2) the nitrogen-doped carbon quantum dot N-CQDs solution of 6 μ L, 8 μ g/mL is dripped to electrode surface, ultrapure water, dry under room temperature;
(3) when electrode is closely dry, drip 3 μ L, 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimides EDC that concentration is 0.3 mol/L and 0.05 mol/L N-hydroxy-succinamide NHS solution is to electrode surface, hatch 30min in 4 DEG C of refrigerators, dry under room temperature;
(4) drip 6 μ L, massfraction be the bovine serum albumin(BSA) BSA solution of 0.5% in electrode surface, with enclosed-electrode nonspecific activity site on the surface, ultrapure water, dries in 4 DEG C of refrigerators; Drip the Tumor specific factor TSGF antigen standard solution of a series of variable concentrations of 6 μ L, 10ng/mL ~ 100 ng/mL in electrode surface, ultrapure water, dry in 4 DEG C of refrigerators;
(5) the detection antibody hatching thing GS-NH of painting 6 μ L, 1 mg/mL is dripped
2@Ab
2solution, in electrode surface, is placed in 4 DEG C of refrigerators and dries, obtained a kind of Tumor specific factor TSGF antigen Electrochemiluminescsensor sensor.
embodiment 2a kind of preparation method of Tumor specific factor TSGF antigen Electrochemiluminescsensor sensor
1. the preparation method of a Tumor specific factor TSGF antigen Electrochemiluminescsensor sensor
(1) by diameter be the glass-carbon electrode Al of 4 mm
2o
3burnishing powder is polished, and ultrapure water cleans up;
(2) the nitrogen-doped carbon quantum dot N-CQDs solution of 6 μ L, 10 μ g/mL is dripped to electrode surface, ultrapure water, dry under room temperature;
(3) when electrode is closely dry, drip 3 μ L, 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimides EDC that concentration is 0.4 mol/L and 0.1 mol/L N-hydroxy-succinamide NHS solution is to electrode surface, hatch 30min in 4 DEG C of refrigerators, dry under room temperature;
(4) drip 6 μ L, bovine serum albumin(BSA) BSA solution that massfraction is 1.0 % in electrode surface, with enclosed-electrode nonspecific activity site on the surface, ultrapure water, dries in 4 DEG C of refrigerators; Drip the Tumor specific factor TSGF antigen standard solution of a series of variable concentrations of 6 μ L, 10ng/mL ~ 100 ng/mL in electrode surface, ultrapure water, dry in 4 DEG C of refrigerators;
(5) the detection antibody hatching thing GS-NH of painting 6 μ L, 2 mg/mL is dripped
2@Ab
2solution, in electrode surface, is placed in 4 DEG C of refrigerators and dries, obtained a kind of Tumor specific factor TSGF antigen Electrochemiluminescsensor sensor.
embodiment 3a kind of preparation method of Tumor specific factor TSGF antigen Electrochemiluminescsensor sensor:
1. the preparation method of a Tumor specific factor TSGF antigen Electrochemiluminescsensor sensor
(1) by diameter be the glass-carbon electrode Al of 4mm
2o
3burnishing powder is polished, and ultrapure water cleans up;
(2) the nitrogen-doped carbon quantum dot N-CQDs solution of 6 μ L, 12 μ g/mL is dripped to electrode surface, ultrapure water, dry under room temperature;
(3) when electrode is closely dry, drip 3 μ L, 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimides EDC that concentration is 0.5 mol/L and 0.2 mol/L N-hydroxy-succinamide NHS solution is to electrode surface, hatch 30min in 4 DEG C of refrigerators, dry under room temperature;
(4) drip 6 μ L, massfraction be the bovine serum albumin(BSA) BSA solution of 2.0% in electrode surface, with enclosed-electrode nonspecific activity site on the surface, ultrapure water, dries in 4 DEG C of refrigerators; Drip the Tumor specific factor TSGF antigen standard solution of a series of variable concentrations of 6 μ L, 10ng/mL ~ 100 ng/mL in electrode surface, ultrapure water, dry in 4 DEG C of refrigerators;
(5) the detection antibody hatching thing GS-NH of painting 6 μ L, 3 mg/mL is dripped
2@Ab
2solution, in electrode surface, is placed in 4 DEG C of refrigerators and dries, obtained a kind of Tumor specific factor TSGF antigen Electrochemiluminescsensor sensor.
embodiment 4the preparation of nitrogen-doped carbon quantum dot N-CQDs
The diethyl pentetic acid DTPA of 0.2 g is dissolved in the ultrapure water of 10 mL, ultrasonic disperse, dissolves at continuing to be heated to 230 DEG C, continues heated solvent evaporation, there is white solid, show the generation of nitrogen-doped carbon quantum dot when white solid becomes brown color; Be dissolved in 20mL ultrapure water by the thick product of gained, 8000 rpmin are centrifugal, and the freeze drying of gained supernatant liquor obtains pure nitrogen-doped carbon quantum dot solid, is ground into powder, obtained nitrogen-doped carbon quantum dot.
embodiment 5the preparation of nitrogen-doped carbon quantum dot N-CQDs
The diethyl pentetic acid DTPA of 0.3 g is dissolved in the ultrapure water of 20mL, ultrasonic disperse, dissolves at continuing to be heated to 230 DEG C, continue heated solvent evaporation, occur white solid, show the generation of nitrogen-doped carbon quantum dot when white solid becomes brown color; Be dissolved in 20mL ultrapure water by the thick product of gained, 8000 rpmin are centrifugal, and the freeze drying of gained supernatant liquor obtains pure nitrogen-doped carbon quantum dot solid, is ground into powder, obtained nitrogen-doped carbon quantum dot.
case study on implementation 6the preparation of nitrogen-doped carbon quantum dot N-CQDs
The diethyl pentetic acid DTPA of 0.4 g is dissolved in the ultrapure water of 30mL, ultrasonic disperse, dissolves at continuing to be heated to 230 DEG C, continue heated solvent evaporation, occur white solid, show the generation of nitrogen-doped carbon quantum dot when white solid becomes brown color; Be dissolved in 20mL ultrapure water by the thick product of gained, 8000 rpmin are centrifugal, and the freeze drying of gained supernatant liquor obtains pure nitrogen-doped carbon quantum dot solid, is ground into powder, obtained nitrogen-doped carbon quantum dot.
embodiment 7a preparation method for Tumor specific factor TSGF antigen Electrochemiluminescsensor sensor, described GS-NH
2@Ab
2detect antibody and incubate compound solution, it is characterized in that, preparation process is as follows:
By the amination Graphene GS-NH of 1mL, 1 mg/mL
2solution, the detection antibody A b of 1mL, 5 μ g/mL
2solution joins in the centrifuge tube of 5 mL, and concussion 12h, makes amination Graphene GS-NH
2with detection antibody A b
2bonding, centrifuging, being distributed to 1mL, pH is in the phosphate buffer solution of 6.5, obtained GS-NH
2@Ab
2detect antibody and incubate compound solution.
embodiment 8a preparation method for Tumor specific factor TSGF antigen Electrochemiluminescsensor sensor, described GS-NH
2@Ab
2detect antibody and incubate compound solution, it is characterized in that, preparation process is as follows:
By the amination Graphene GS-NH of 1mL, 2 mg/mL
2solution, the detection antibody A b of 1mL, 10 μ g/mL
2solution joins in the centrifuge tube of 5 mL, and concussion 12h, makes amination Graphene GS-NH
2with detection antibody A b
2bonding, centrifuging, being distributed to 1mL, pH is in the phosphate buffer solution of 7.5, obtained GS-NH
2@Ab
2detect antibody and incubate compound solution.
embodiment 9a preparation method for Tumor specific factor TSGF antigen Electrochemiluminescsensor sensor, described GS-NH
2@Ab
2detect antibody and incubate compound solution, it is characterized in that, preparation process is as follows:
By the amination Graphene GS-NH of 1mL, 3 mg/mL
2solution, the detection antibody A b of 1mL, 15 μ g/mL
2solution joins in the centrifuge tube of 5 mL, and concussion 12h, makes amination Graphene GS-NH
2with detection antibody A b
2bonding, centrifuging, being distributed to 1mL, pH is in the phosphate buffer solution of 8.5, obtained GS-NH
2@Ab
2detect antibody and incubate compound solution.
embodiment 10an Electrochemiluminescsensor sensor for nitrogen-doped carbon quantum dot, for the detection of Tumor specific factor TSGF antigen, detecting step is as follows:.
(1) three-electrode system of electrochemical workstation is used to test, Ag/AgCl electrode is as contrast electrode, platinum electrode is to electrode, prepared electrochemical luminous sensor is working electrode, electrochemical workstation and chemiluminescence detector are linked together the high pressure of photomultiplier is set to 650 V, scanning voltage is set to-2 ~ 0 V, sweep speed 0.1 V/s;
(2) 10 mL, pH 7.5 containing 80 mmol/L potassium persulfates PBS buffer solution in, by the Tumor specific factor TSGF antigen standard solution of electrogenerated chemiluminescence systems axiol-ogy to a series of variable concentrations of 0.01 ng/mL ~ 100 ng/mL, the electrochemiluminescence signal intensity produced, drawing curve;
(3) replaced by testing sample solution Tumor specific factor TSGF antigen standard solution to detect, recording the range of linearity is 0.01 ng/mL ~ 100 ng/mL, detects and is limited to 3.3 pg/mL.
Claims (5)
1. a preparation method for Tumor specific factor TSGF antigen Electrochemiluminescsensor sensor, it is characterized in that, step is as follows:
(1) by diameter be the glass-carbon electrode Al of 4mm
2o
3burnishing powder is polished, and ultrapure water cleans up;
(2) the nitrogen-doped carbon quantum dot N-CQDs solution of 6 μ L, 8 ~ 12 μ g/mL is dripped to electrode surface, ultrapure water, dry under room temperature;
(3) when electrode is closely dry, drip 3 μ L, 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimides EDC that concentration is 0.3 ~ 0.5 mol/L and 0.05 ~ 0.2 mol/L N-hydroxy-succinamide NHS solution is to electrode surface, hatch 30min in 4 DEG C of refrigerators, dry under room temperature;
(4) drip 6 μ L, massfraction be the bovine serum albumin(BSA) BSA solution of 0.5 ~ 2.0% in electrode surface, with enclosed-electrode nonspecific activity site on the surface, ultrapure water, dries in 4 DEG C of refrigerators; Drip the Tumor specific factor TSGF antigen standard solution of a series of variable concentrations of 6 μ L, 10ng/mL ~ 100 ng/mL in electrode surface, ultrapure water, dry in 4 DEG C of refrigerators;
(5) the detection antibody hatching thing GS-NH of painting 6 μ L, 1 ~ 3 mg/mL is dripped
2@Ab
2solution, in electrode surface, is placed in 4 DEG C of refrigerators and dries, obtained a kind of Tumor specific factor TSGF antigen Electrochemiluminescsensor sensor.
2. the preparation method of a kind of Tumor specific factor TSGF antigen Electrochemiluminescsensor sensor as claimed in claim 1, described nitrogen-doped carbon quantum dot N-CQDs, it is characterized in that, preparation process is as follows:
The diethyl pentetic acid DTPA of 0.2 ~ 0.4 g is dissolved in the ultrapure water of 10 ~ 30mL, ultrasonic disperse, dissolves at continuing to be heated to 230 DEG C, continues heated solvent evaporation, there is white solid, show the generation of nitrogen-doped carbon quantum dot N-CQDs when white solid becomes brown color; Be dissolved in 20mL ultrapure water by the thick product of gained, 8000 rpmin are centrifugal, and the freeze drying of gained supernatant liquor obtains pure nitrogen-doped carbon quantum dot N-CQDs solid, is ground into powder, obtained nitrogen-doped carbon quantum dot N-CQDs.
3. the preparation method of a kind of Tumor specific factor TSGF antigen Electrochemiluminescsensor sensor as claimed in claim 1, described detection antibody hatching thing GS-NH
2@Ab
2solution, is characterized in that, preparation process is as follows:
By the amination Graphene GS-NH of 1mL, 1 ~ 3 mg/mL
2solution, the detection antibody A b of 1mL, 5 ~ 10 μ g/mL
2solution joins in the centrifuge tube of 5 mL, and concussion 12h, makes amination Graphene GS-NH
2with detection antibody A b
2bonding, centrifuging, being distributed to 1mL, pH is in the phosphate buffer solution of 6.5 ~ 8.5, and the obtained antibody that detects hatches thing GS-NH
2@Ab
2solution.
4. a kind of Electrochemiluminescsensor sensor of nitrogen-doped carbon quantum dot prepared of preparation method as claimed in claim 1, for the detection of Tumor specific factor TSGF antigen, detecting step is as follows:
(1) three-electrode system of electrochemical workstation is used to test, Ag/AgCl electrode is as contrast electrode, platinum electrode is to electrode, prepared Electrochemiluminescsensor sensor is working electrode, electrochemical workstation and chemiluminescence detector are linked together the high pressure of photomultiplier is set to 650 V, scanning voltage is set to-2 ~ 0 V, sweep speed 0.1 V/s;
(2) 10 mL, pH 7.5 containing 80 mmol/L potassium persulfates PBS buffer solution in, by the Tumor specific factor TSGF antigen standard solution of electrogenerated chemiluminescence systems axiol-ogy to a series of variable concentrations of 0.01 ng/mL ~ 100 ng/mL, the electrochemiluminescence signal intensity produced, drawing curve;
(3) Tumor specific factor TSGF antigen standard solution is replaced by testing sample solution to detect.
5. the preparation method of a kind of Tumor specific factor TSGF antigen Electrochemiluminescsensor sensor as claimed in claim 1, described tumor markers is a kind of Tumor specific factor TSGF antigen.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105136879A (en) * | 2015-08-20 | 2015-12-09 | 济南大学 | Preparation method and application of electrogenerated chemiluminescence sensor based on graphene quantum dots |
| CN105675679A (en) * | 2016-03-04 | 2016-06-15 | 济南大学 | Preparation and application of ZnO-NCQDs (Nitrogen-doped Carbon Quantum Dots) DNA photoelectric sensor |
| CN105717098A (en) * | 2016-02-25 | 2016-06-29 | 济南大学 | Preparation method and application of electrochemiluminescent bisphenol A biosensor based on nitrogen-doped titanium dioxide nanosheet |
| CN105806922A (en) * | 2016-03-16 | 2016-07-27 | 济南大学 | Preparation method of parathion-methyl immunosensor based on combination of two electrochemical methods and application |
| CN106053444A (en) * | 2016-04-28 | 2016-10-26 | 安徽伊普诺康生物技术股份有限公司 | A kit for measuring a specific growth factor and a preparing method thereof |
| CN107741498A (en) * | 2017-10-31 | 2018-02-27 | 宁波卫生职业技术学院 | A kind of preparation method for being used to detect tumor markers immunosensor |
| CN110016339A (en) * | 2019-05-07 | 2019-07-16 | 山西大学 | A kind of room temperature phosphorimetry carbon quantum dot and its preparation method and application that daylight can excite |
| CN111007136A (en) * | 2019-12-28 | 2020-04-14 | 杭州育源生命科技有限公司 | Preparation of immunosensor and method for FSH antibody detection |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102967706A (en) * | 2012-11-21 | 2013-03-13 | 济南大学 | Preparation method and application of flow injection chemiluminiscence immuno sensor for detecting tumor marker |
| CN104297480A (en) * | 2014-09-26 | 2015-01-21 | 济南大学 | Preparation method and application of sandwich type immunosensor for prostate specific antigens |
| CN204287194U (en) * | 2014-12-23 | 2015-04-22 | 苏州大学 | Based on the electrochemiluminescence immunosensor of carbon quantum dot-polyamide-amide dendrimer |
-
2015
- 2015-05-13 CN CN201510241947.6A patent/CN104833806B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102967706A (en) * | 2012-11-21 | 2013-03-13 | 济南大学 | Preparation method and application of flow injection chemiluminiscence immuno sensor for detecting tumor marker |
| CN104297480A (en) * | 2014-09-26 | 2015-01-21 | 济南大学 | Preparation method and application of sandwich type immunosensor for prostate specific antigens |
| CN204287194U (en) * | 2014-12-23 | 2015-04-22 | 苏州大学 | Based on the electrochemiluminescence immunosensor of carbon quantum dot-polyamide-amide dendrimer |
Non-Patent Citations (3)
| Title |
|---|
| HAIXIN CHANG等: "Graphene fluorescence resonance energy transfer aptasensor for the thrombin detection.", 《ANAL CHEM》 * |
| TONGQIAN HAN等: "Eco-friendly synthesis of electrochemiluminescent nitrogen-doped carbon quantum dots from diethylene triamine pentacetate and their application for protein detection.", 《CARBON》 * |
| W.WEI等: "Electrochemiluminescent detection of Mucin 1 protein and MCF-7 cancer cells based on the resonance energy transfer.", 《ANALYST》 * |
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| CN105717098B (en) * | 2016-02-25 | 2018-03-27 | 济南大学 | A kind of preparation method and application of the electrogenerated chemiluminescence bisphenol-A biology sensor based on nitrogen-doped titanium dioxide nanometer sheet |
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| CN105806922B (en) * | 2016-03-16 | 2018-05-08 | 济南大学 | A kind of preparation method and application based on parathion-methyl immunosensor associated with two kinds of electrochemical methods |
| CN106053444A (en) * | 2016-04-28 | 2016-10-26 | 安徽伊普诺康生物技术股份有限公司 | A kit for measuring a specific growth factor and a preparing method thereof |
| CN107741498A (en) * | 2017-10-31 | 2018-02-27 | 宁波卫生职业技术学院 | A kind of preparation method for being used to detect tumor markers immunosensor |
| CN110016339A (en) * | 2019-05-07 | 2019-07-16 | 山西大学 | A kind of room temperature phosphorimetry carbon quantum dot and its preparation method and application that daylight can excite |
| CN110016339B (en) * | 2019-05-07 | 2021-07-02 | 山西大学 | Sunlight-excitable room-temperature phosphorescent carbon quantum dot and preparation method and application thereof |
| CN111007136A (en) * | 2019-12-28 | 2020-04-14 | 杭州育源生命科技有限公司 | Preparation of immunosensor and method for FSH antibody detection |
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