CN110455786B - Based on CeO2@SnS2Preparation method of luminol-promoted electrochemiluminescence sensor - Google Patents
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Abstract
The invention relates to a CeO-based material2@SnS2A preparation method of a luminol-promoted electrochemiluminescence sensor belongs to the field of novel nano materials and the technical field of biosensing; the invention is based on ECL technology, ferritin covalent cross-linking luminol (Ft-luminol) is used as a signal source for the first time, and CeO is used2@SnS2As accelerator, use is made of CeO2@SnS2The excellent synergistic catalytic action of luminol can effectively amplify detection signals, the preparation method of the biosensor is simple in preparation, low in cost, low in reaction energy consumption, green and environment-friendly, and is applied to the detection of an actual sample of procalcitonin, the detection limit is as low as 1.6 fg/mL, the linear range is as wide as 5 fg/mL-100 ng/mL, the sensitivity is high, the reproducibility is good, and the potential application value is high.
Description
Technical Field
The invention belongs to the field of novel nano materials and the technical field of biosensing
Background
As a research hotspot arising from the interdigitation of various disciplines such as biology, chemistry, medicine, electronic technology and the like, the Electrochemiluminescence (ECL) immunoassay technology is the organic combination of electrochemistry, chemiluminescence and immunoassay technology, has the advantages of low cost, good selectivity, high sensitivity, high analysis speed, easy automation, miniaturization, integration and the like, and is widely applied to the fields of disease marker analysis, food safety analysis, environmental pollution analysis and the like.
The procalcitonin is used as a disease marker for diagnosing sepsis and identifying severe bacterial infection, and the sensitivity and the specificity of sepsis diagnosis can be obviously improved by sensitive detection of the procalcitonin. At present, electrochemical biosensors are mainly used for detecting procalcitonin, but the method has the defects of poor reproducibility and low sensitivity. The electrochemiluminescence technology combines the advantages of the chemiluminescence technology and the electrochemistry technology, and has the characteristics of easy operation, strong controllability, high response speed and the like, the sensitivity is higher than that of electrochemistry, and the detection limit is lower.
Disclosure of Invention
One of the technical tasks of the invention is to make up the defects of the prior detection technology and provide a reagent prepared from CeO2@SnS2The immune sensing technology for promoting luminol electrochemiluminescence has the advantages of simple operation, low cost, rapid signal response, greatly shortened detection time, and time and labor saving.
The second technical task of the invention is to provide the application of the biosensor, the sensor can rapidly detect procalcitonin, and has the advantages of high sensitivity, strong specificity and good reproducibility, the detection limit is 1.6 fg/mL, and the linear range is 5 fg/mL-50 ng/mL.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
based on CeO2@SnS2The preparation method of the luminol-promoted electrochemiluminescence sensor comprises the following steps:
(1) polishing glassy carbon electrodes with the diameter of 4 mm by using alumina polishing powder with the diameter of 1.0 micron, 0.3 micron and 0.05 micron in sequence, and washing the polished glassy carbon electrodes with ultrapure water;
(2) dripping 6 mu L of CeO with the concentration of 2-4 mg/mL on the surface of a glassy carbon electrode2@SnS2The material solution is used as a sensing substrate and is placed at 37 ℃ for airing;
(3) dripping 6 mu L of antibody-labeled luminol-ferritin (Ft-luminol) solution with the concentration of 2-4 mg/mL, washing the surface of the electrode by phosphate buffer solution PBS with the pH of 7.4, and placing the electrode at 4 ℃ for airing;
(4) dropwise adding 3 mu L of bovine serum albumin solution with the mass fraction of 1-3% to seal non-specific active sites on the surface of the electrode, washing the surface of the electrode by phosphate buffer solution PBS with the pH of 7.4, and placing the electrode at 4 ℃ for airing;
(5) dripping 6 mu L of procalcitonin standard solution or procalcitonin solution with unknown concentration, incubating for 0.5-2 h at 37 ℃, washing the surface of the electrode by phosphate buffer solution PBS with pH 7.4, placing the electrode at 4 ℃, airing, and finishing the construction of the sensor.
Based on CeO2@SnS2Preparation method of luminol-promoted electrochemiluminescence sensor and CeO2@SnS2The material solution is prepared by the following steps:
firstly, 0.01-0.03 mol of tin tetrachloride pentahydrate, 0.001-0.003 mol of cerium nitrate hexahydrate, 0.04-0.06 mol of thioacetamide and 0.4-0.6 g of hexadecyltrimethylammonium bromide are completely dissolved in a mixture of ethanol and distilled water, stirred for 2 hours at 80 ℃, the obtained product is transferred into a 200 mL high-temperature reaction kettle with a polytetrafluoroethylene lining, the temperature is kept at 180 ℃, the reaction lasts for 24-48 hours, and then the mixture is cooled to room temperature. Centrifugally washing the obtained solid for multiple times, drying the solid in a vacuum drying oven at 40-80 ℃ for 12-36 hours, and sintering the solid at 500 ℃ for 3-6 hours under the protection of argon to obtain CeO2@SnS2Dissolving 2-4 mg of product in 1 mL of ultrapure water to prepare CeO with the concentration of 2-4 mg/mL2@SnS2And (3) solution.
Based on CeO2@SnS2Promoting the preparation method of the luminol electrochemiluminescence sensor,the antibody-labeled luminol-ferritin (Ft-luminol) solution is prepared by the following steps:
mixing 3-5 mL of ferritin solution with the concentration of 5 mug/mL and 0.5-2.5 mL of luminol solution with the concentration of 10 mmol/L, then adding 3.5-5.5 mL of deionized water for dilution, then adding 50-150 uL of glutaraldehyde solution with the mass fraction of 50% into the mixed solution, continuously stirring for 2 hours to obtain pale yellow luminol-ferritin solution, continuously adding 100-300 uL of procalcitonin antibody solution with the concentration of 10 mg/mL, oscillating and incubating for 6-18 hours at 4 ℃, centrifuging, dispersing into 1 mL of phosphate buffer solution with the pH of 7.4 to obtain antibody-labeled luminol-ferritin solution, and storing at 4 ℃ for later use.
Based on CeO2@SnS2The preparation method of the luminol electrochemiluminescence sensor is promoted, and the electrochemiluminescence sensor prepared based on the method is used for detecting the concentration of procalcitonin.
The electrochemiluminescence sensor is used for detecting the concentration of procalcitonin, and the operation steps are as follows:
(1) setting parameters: the high voltage of a photomultiplier of the ultra-weak electrochemiluminescence instrument is set to be 800V, the cyclic volt-ampere scanning potential range of the electrochemical workstation is set to be 0-0.6V, and the scanning rate is set to be 0.1V/s;
(2) and (3) testing: taking a silver/silver chloride electrode as a reference electrode, a platinum wire electrode as a counter electrode, taking the sensor prepared by the method as a working electrode, carrying out electrochemiluminescence test in 10 mL of phosphate buffer solution containing 45-75 mmol/L of hydrogen peroxide to obtain corresponding electrochemiluminescence signal intensity when hatching procalcitonin with different concentrations, and drawing a working curve, wherein the detection limit is 1.6 fg/mL, and the linear range is 5 fg/mL-50 ng/mL;
(3) and testing the electrochemiluminescence sensor for incubating the procalcitonin actual sample with unknown concentration to obtain corresponding signal intensity, and calculating according to the working curve to obtain the procalcitonin concentration in the actual sample.
Advantageous results of the invention
（1）Based on CeO for the first time2@SnS2The principle of promoting luminol electrochemiluminescence provides a novel and reliable immunosensing technology. Since CeO2And SnS2Each has good electrocatalysis performance, and the CeO is obtained by successfully compounding the two through a one-pot hydrothermal synthesis method2@SnS2Using CeO2And SnS2With satisfactory synergistic effect therebetween, CeO2@SnS2The hydrogen peroxide can be efficiently catalyzed and decomposed to generate superoxide anions and hydroxyl free radicals, so that sufficient co-reaction free radicals are provided for ECL excitation of luminol, and the effective amplification of ECL signals is realized;
(2) the method solves the problems of complex operation, low sensitivity and poor reproducibility of the existing electrochemical detection technology, is applied to the sample detection of procalcitonin, has the detection limit of 1.6 fg/mL and the linear range of 5 fg/mL-50 ng/mL, and has the advantages of high response speed, high sensitivity, good reproducibility, simple preparation, low cost and environmental protection.
Detailed Description
The invention will now be further illustrated by reference to specific examples, which are intended to be illustrative only and not to limit the scope of the invention.
Example 1A composition based on CeO2@SnS2The preparation method of the luminol-promoted electrochemiluminescence sensor comprises the following steps:
(1) polishing glassy carbon electrodes with the diameter of 4 mm by using alumina polishing powder with the diameter of 1.0 micron, 0.3 micron and 0.05 micron in sequence, and washing the polished glassy carbon electrodes with ultrapure water;
(2) dropping 6 mu L of CeO with the concentration of 2 mg/mL on the surface of the glassy carbon electrode2@SnS2The material solution is used as a sensing substrate and is placed at 37 ℃ for airing;
(3) dripping 6 mu L of antibody-labeled luminol-ferritin (Ft-luminol) solution with the concentration of 2 mg/mL, washing the surface of the electrode by phosphate buffer solution PBS with the pH value of 7.4, and placing the electrode at 4 ℃ for airing;
(4) dropwise adding 3 mu L of bovine serum albumin solution with the mass fraction of 1% to seal the nonspecific active sites on the surface of the electrode, washing the surface of the electrode with phosphate buffer solution PBS with the pH of 7.4, and placing the electrode at 4 ℃ for airing;
(5) dripping 6 mu L of procalcitonin standard solution or procalcitonin solution with unknown concentration, incubating at 37 ℃ for 0.5 h, washing the surface of the electrode with phosphate buffer solution PBS with pH 7.4, placing the electrode at 4 ℃ for airing, and finishing the construction of the sensor.
Example 2A composition based on CeO2@SnS2The preparation method of the luminol-promoted electrochemiluminescence sensor comprises the following steps:
(1) polishing glassy carbon electrodes with the diameter of 4 mm by using alumina polishing powder with the diameter of 1.0 micron, 0.3 micron and 0.05 micron in sequence, and washing the polished glassy carbon electrodes with ultrapure water;
(2) dropping 6 mu L of CeO with the concentration of 3 mg/mL on the surface of the glassy carbon electrode2@SnS2The material solution is used as a sensing substrate and is placed at 37 ℃ for airing;
(3) dripping 6 mu L of antibody-labeled luminol-ferritin (Ft-luminol) solution with the concentration of 3 mg/mL, washing the surface of the electrode by phosphate buffer solution PBS with the pH value of 7.4, and placing the electrode at 4 ℃ for airing;
(4) dropwise adding 3 mu L of 2% bovine serum albumin solution to seal the nonspecific active sites on the surface of the electrode, washing the surface of the electrode with phosphate buffer solution PBS (pH 7.4), and airing at 4 ℃;
(5) dripping 6 mu L of procalcitonin standard solution or procalcitonin solution with unknown concentration, incubating for 1 h at 37 ℃, washing the surface of the electrode by phosphate buffer solution PBS with pH 7.4, placing the electrode at 4 ℃, airing, and finishing the construction of the sensor.
Example 3A composition based on CeO2@SnS2The preparation method of the luminol-promoted electrochemiluminescence sensor comprises the following steps:
(1) polishing glassy carbon electrodes with the diameter of 4 mm by using alumina polishing powder with the diameter of 1.0 micron, 0.3 micron and 0.05 micron in sequence, and washing the polished glassy carbon electrodes with ultrapure water;
(2) dropping 6 mu L of CeO with the concentration of 4 mg/mL on the surface of the glassy carbon electrode2@SnS2The material solution is used as a sensing substrate and is placed at 37 ℃ for airing;
(3) dripping 6 mu L of antibody-labeled luminol-ferritin (Ft-luminol) solution with the concentration of 4 mg/mL, washing the surface of the electrode by phosphate buffer solution PBS with the pH value of 7.4, and placing the electrode at 4 ℃ for airing;
(4) dropwise adding 3 mu L of bovine serum albumin solution with the mass fraction of 3% to seal the nonspecific active sites on the surface of the electrode, washing the surface of the electrode with phosphate buffer solution PBS with the pH of 7.4, and placing the electrode at 4 ℃ for airing;
(5) dripping 6 mu L of procalcitonin standard solution or procalcitonin solution with unknown concentration, incubating for 2 h at 37 ℃, washing the surface of the electrode by phosphate buffer solution PBS with pH 7.4, placing the electrode at 4 ℃, airing, and finishing the construction of the sensor.
Example 4. the CeO2@SnS2The material solution is prepared by the following steps:
first, a mixture of 0.01 mol of tin tetrachloride pentahydrate, 0.001 mol of cerium nitrate hexahydrate, 0.04 mol of thioacetamide, 0.4 g of cetyltrimethylammonium bromide completely dissolved in ethanol and distilled water was stirred at 80 ℃ for 2 hours, and the resultant was transferred to a 200 mL polytetrafluoroethylene-lined high-temperature reaction vessel, the temperature was maintained at 180 ℃ for 24 hours, followed by cooling to room temperature. Centrifugally washing the obtained solid for multiple times, drying the solid in a vacuum drying oven at 40 ℃ for 12 hours, and then sintering the solid for 3 hours at 500 ℃ under the protection of argon to obtain CeO2@SnS2Dissolving 2 mg of the product in 1 mL of ultrapure water to prepare CeO with the concentration of 2 mg/mL2@SnS2And (3) solution.
Example 5. the CeO2@SnS2A material solution prepared by the steps of:
first, 0.02 mol of tin tetrachloride pentahydrate, 0.002 mol of cerium nitrate hexahydrate, 0.05 mol of thioacetamide, 0.5 g of cetyltrimethylammonium bromide completely dissolved in a mixture of ethanol and distilled water were stirred at 80 ℃ for 2 hours, the resultant was transferred to a 200 mL polytetrafluoroethylene-lined high-temperature reaction vessel, the temperature was maintained at 180 ℃ for 36 hours, and subsequentlyAnd cooling to room temperature. Centrifugally washing the obtained solid for multiple times, drying the solid in a vacuum drying oven at 60 ℃ for 24 hours, and then sintering the solid for 4 hours at 500 ℃ under the protection of argon to obtain CeO2@SnS2Dissolving 3 mg of the product in 1 mL of ultrapure water to prepare CeO with the concentration of 3 mg/mL2@SnS2And (3) solution.
Example 6. the CeO2@SnS2The material solution is prepared by the following steps:
first, a mixture of 0.03 mol of tin tetrachloride pentahydrate, 0.003 mol of cerium nitrate hexahydrate, 0.06 mol of thioacetamide, 0.6 g of cetyltrimethylammonium bromide completely dissolved in ethanol and distilled water was stirred at 80 ℃ for 2 hours, and the resultant was transferred to a 200 mL polytetrafluoroethylene-lined high-temperature reaction vessel, the temperature was maintained at 180 ℃ for 48 hours, followed by cooling to room temperature. Centrifugally washing the obtained solid for multiple times, drying the solid in a vacuum drying oven at 80 ℃ for 36 hours, and then sintering the solid for 6 hours at 500 ℃ under the protection of argon to obtain CeO2@SnS2Dissolving 4 mg of the product in 1 mL of ultrapure water to prepare CeO with the concentration of 4 mg/mL2@SnS2And (3) solution.
Example 7 antibody-labeled luminol-ferritin (Ft-luminol) solution, prepared by the following steps:
mixing 3 mL of ferritin solution with the concentration of 5 mug/mL with 0.5 mL of luminol solution with the concentration of 10 mmol/L, adding 3.5 mL of deionized water for dilution, adding 50 uL of glutaraldehyde solution with the mass fraction of 50% into the mixed solution, continuously stirring for 2 hours to obtain pale yellow luminol-ferritin solution, continuously adding 100 uL of procalcitonin antibody solution with the concentration of 10 mg/mL, oscillating and incubating for 6 hours at 4 ℃, centrifuging, dispersing into 1 mL of phosphate buffer solution with the pH of 7.4 to obtain antibody-labeled luminol-ferritin solution, and storing at 4 ℃ for later use.
Example 8 antibody-labeled luminol-ferritin (Ft-luminol) solution, prepared by the following steps:
mixing 4 mL of ferritin solution with the concentration of 5 mug/mL with 1.5 mL of luminol solution with the concentration of 10 mmol/L, adding 4.5 mL of deionized water for dilution, then adding 100 uL of glutaraldehyde solution with the mass fraction of 50% into the mixed solution, continuously stirring for 2 hours to obtain pale yellow luminol-ferritin solution, continuously adding 200 uL of procalcitonin antibody solution with the concentration of 10 mg/mL, oscillating and incubating for 12 hours at 4 ℃, centrifuging, dispersing into 1 mL of phosphate buffer solution with the pH of 7.4 to obtain antibody-labeled luminol-ferritin solution, and storing at 4 ℃ for later use.
Example 9 antibody-labeled luminol-ferritin (Ft-luminol) solution, prepared by the following steps:
mixing 5 mL of ferritin solution with the concentration of 5 mug/mL with 2.5 mL of luminol solution with the concentration of 10 mmol/L, adding 5.5 mL of deionized water for dilution, then adding 150 uL of glutaraldehyde solution with the mass fraction of 50% into the mixed solution, continuously stirring for 2 hours to obtain pale yellow luminol-ferritin solution, continuously adding 300 uL of procalcitonin antibody solution with the concentration of 10 mg/mL, oscillating and incubating for 18 hours at 4 ℃, centrifuging, dispersing into 1 mL of phosphate buffer solution with the pH of 7.4 to obtain antibody-labeled luminol-ferritin solution, and storing at 4 ℃ for later use.
Example 10 the sensor is used for the detection of procalcitonin concentration, and the operation steps are as follows:
(1) setting parameters: the high voltage of a photomultiplier of the ultra-weak electrochemiluminescence instrument is set to be 800V, the cyclic volt-ampere scanning potential range of the electrochemical workstation is set to be 0-0.6V, and the scanning rate is set to be 0.1V/s;
(2) and (3) testing: taking a silver/silver chloride electrode as a reference electrode, a platinum wire electrode as a counter electrode, taking the sensor prepared by the method as a working electrode, carrying out electrochemiluminescence test in 10 mL of phosphate buffer solution containing 45 mmol/L hydrogen peroxide to obtain corresponding electrochemiluminescence signal intensity when hatching procalcitonin with different concentrations, and drawing a working curve, wherein the detection limit is 1.6 fg/mL, and the linear range is 5 fg/mL-50 ng/mL;
(3) and testing the electrochemiluminescence sensor for incubating the procalcitonin actual sample with unknown concentration to obtain corresponding signal intensity, and calculating according to the working curve to obtain the procalcitonin concentration in the actual sample.
Example 11. the sensor was used for the detection of procalcitonin concentration, and the procedure was as follows:
(1) setting parameters: the high voltage of a photomultiplier of the ultra-weak electrochemiluminescence instrument is set to be 800V, the cyclic volt-ampere scanning potential range of the electrochemical workstation is set to be 0-0.6V, and the scanning rate is set to be 0.1V/s;
(2) and (3) testing: taking a silver/silver chloride electrode as a reference electrode, a platinum wire electrode as a counter electrode, taking the sensor prepared by the method as a working electrode, carrying out electrochemiluminescence test in 10 mL of phosphate buffer solution containing 55 mmol/L of hydrogen peroxide to obtain corresponding electrochemiluminescence signal intensity when hatching procalcitonin with different concentrations, and drawing a working curve, wherein the detection limit is 1.6 fg/mL, and the linear range is 5 fg/mL-50 ng/mL;
(3) and testing the electrochemiluminescence sensor for incubating the procalcitonin actual sample with unknown concentration to obtain corresponding signal intensity, and calculating according to the working curve to obtain the procalcitonin concentration in the actual sample.
Example 12. the sensor was used for the detection of procalcitonin concentration, the procedure was as follows:
(1) setting parameters: the high voltage of a photomultiplier of the ultra-weak electrochemiluminescence instrument is set to be 800V, the cyclic volt-ampere scanning potential range of the electrochemical workstation is set to be 0-0.6V, and the scanning rate is set to be 0.1V/s;
(2) and (3) testing: taking a silver/silver chloride electrode as a reference electrode, a platinum wire electrode as a counter electrode, taking the sensor prepared by the method as a working electrode, carrying out electrochemiluminescence test in 10 mL of phosphate buffer solution containing 75 mmol/L hydrogen peroxide to obtain corresponding electrochemiluminescence signal intensity when hatching procalcitonin with different concentrations, and drawing a working curve, wherein the detection limit is 1.6 fg/mL, and the linear range is 5 fg/mL-50 ng/mL;
(3) and testing the electrochemiluminescence sensor for incubating the procalcitonin actual sample with unknown concentration to obtain corresponding signal intensity, and calculating according to the working curve to obtain the procalcitonin concentration in the actual sample.
Claims (5)
1. Based on CeO2@SnS2The preparation method of the luminol-promoted electrochemiluminescence sensor is characterized by comprising the following steps:
(1) polishing glassy carbon electrodes with the diameter of 4 mm by using alumina polishing powder with the diameter of 1.0 micron, 0.3 micron and 0.05 micron in sequence, and washing the polished glassy carbon electrodes with ultrapure water;
(2) dripping 6 mu L of CeO with the concentration of 2-4 mg/mL on the surface of a glassy carbon electrode2@SnS2The material solution is used as a sensing substrate and is placed at 37 ℃ for airing;
(3) dripping 6 mu L of antibody-labeled luminol-ferritin (Ft-luminol) solution with the concentration of 2-4 mg/mL, washing the surface of the electrode by phosphate buffer solution PBS with the pH of 7.4, and placing the electrode at 4 ℃ for airing;
(4) dropwise adding 3 mu L of bovine serum albumin solution with the mass fraction of 1-3% to seal non-specific active sites on the surface of the electrode, washing the surface of the electrode by phosphate buffer solution PBS with the pH of 7.4, and placing the electrode at 4 ℃ for airing;
(5) dripping 6 mu L of procalcitonin standard solution or procalcitonin solution with unknown concentration, incubating for 0.5-2 h at 37 ℃, washing the surface of the electrode by phosphate buffer solution PBS with pH 7.4, placing the electrode at 4 ℃, airing, and finishing the construction of the sensor.
2. CeO-based alloy according to claim 12@SnS2The preparation method of the luminol-promoted electrochemiluminescence sensor is characterized in that the CeO2@SnS2The material solution is prepared by the following steps:
firstly, 0.01 to 0.03 mol of tin tetrachloride pentahydrate, 0.001 to 0.003 mol of cerium nitrate hexahydrate, 0.04 to 0.06 mol of thioacetamide and 0.4 to 0.6 g of cetyltrimethylammonium bromide are completely dissolved in a mixture of ethanol and distilled water, stirred for 2 hours at 80 ℃, and the obtained product is transferred into a 200 mL high-temperature reaction kettle with a polytetrafluoroethylene lining, and the temperature is kept at 180 DEG CReacting for 24-48 h, cooling to room temperature, centrifugally washing the obtained solid for multiple times, drying in a vacuum drying oven at 40-80 ℃ for 12-36 h, and sintering at 500 ℃ for 3-6 h under the protection of argon to obtain CeO2@SnS2Dissolving 2-4 mg of product in 1 mL of ultrapure water to prepare CeO with the concentration of 2-4 mg/mL2@SnS2And (3) solution.
3. CeO-based alloy according to claim 12@SnS2The preparation method of the luminol-promoted electrochemiluminescence sensor is characterized in that the luminol-ferritin (Ft-luminol) solution labeled by the antibody is prepared according to the following steps:
mixing 3-5 mL of ferritin solution with the concentration of 5 mug/mL and 0.5-2.5 mL of luminol solution with the concentration of 10 mmol/L, then adding 3.5-5.5 mL of deionized water for dilution, then adding 50-150 uL of glutaraldehyde solution with the mass fraction of 50% into the mixed solution, continuously stirring for 2 hours to obtain pale yellow luminol-ferritin solution, continuously adding 100-300 uL of procalcitonin antibody solution with the concentration of 10 mg/mL, oscillating and incubating for 6-18 hours at 4 ℃, centrifuging, dispersing into 1 mL of phosphate buffer solution with the pH of 7.4 to obtain antibody-labeled luminol-ferritin solution, and storing at 4 ℃ for later use.
4. The electrochemiluminescence sensor prepared by the preparation method of claim 1 is used for detecting the concentration of procalcitonin.
5. The use of an electrochemiluminescence sensor according to claim 4 for the detection of procalcitonin concentration, characterized by the following operating steps:
(1) setting parameters: the high voltage of a photomultiplier of the ultra-weak electrochemiluminescence instrument is set to be 800V, the cyclic volt-ampere scanning potential range of the electrochemical workstation is set to be 0-0.6V, and the scanning rate is set to be 0.1V/s;
(2) and (3) testing: taking a silver/silver chloride electrode as a reference electrode, a platinum wire electrode as a counter electrode, taking the sensor prepared by the method as a working electrode, carrying out electrochemiluminescence test in 10 mL of phosphate buffer solution containing 45-75 mmol/L of hydrogen peroxide to obtain corresponding electrochemiluminescence signal intensity when hatching procalcitonin with different concentrations, and drawing a working curve, wherein the detection limit is 1.6 fg/mL, and the linear range is 5 fg/mL-50 ng/mL;
(3) and testing the electrochemiluminescence sensor for incubating the procalcitonin actual sample with unknown concentration to obtain corresponding signal intensity, and calculating according to the working curve to obtain the procalcitonin concentration in the actual sample.
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| CN111766289B (en) * | 2020-06-22 | 2022-08-30 | 济南大学 | Oxygen-enriched vacancy CeO 2 Preparation method of electrochemiluminescence immunosensor |
| CN111766288B (en) * | 2020-06-22 | 2022-10-28 | 济南大学 | Based on oxygen boosting vacancy NiCo 2 O 4 Preparation method of electrochemiluminescence sensor |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104880456A (en) * | 2015-05-22 | 2015-09-02 | 济南大学 | Preparation method and application of electrochemiluminescence immunosensor constructed on basis of GO/MWCNTs-COOH/Au @ CeO2 |
| CN106404756A (en) * | 2016-09-05 | 2017-02-15 | 济南大学 | Preparation method and application of electrochemiluminescence sensor based on graphene/Fe3O4@Au/CeO2/TiO2 |
| CN109507174A (en) * | 2019-01-16 | 2019-03-22 | 济南大学 | Preparation based on the compound ZnO nanoparticle quenching Particles in Electrochemiluminescofce ofce Luminol sensor of curcumin |
| CN109668874A (en) * | 2018-12-28 | 2019-04-23 | 济南大学 | A kind of preparation method of the electrochemical luminescence immunosensor of different luminol functionalization MOFs detection beta-amyloid protein |
-
2019
- 2019-08-26 CN CN201910787978.XA patent/CN110455786B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104880456A (en) * | 2015-05-22 | 2015-09-02 | 济南大学 | Preparation method and application of electrochemiluminescence immunosensor constructed on basis of GO/MWCNTs-COOH/Au @ CeO2 |
| CN106404756A (en) * | 2016-09-05 | 2017-02-15 | 济南大学 | Preparation method and application of electrochemiluminescence sensor based on graphene/Fe3O4@Au/CeO2/TiO2 |
| CN109668874A (en) * | 2018-12-28 | 2019-04-23 | 济南大学 | A kind of preparation method of the electrochemical luminescence immunosensor of different luminol functionalization MOFs detection beta-amyloid protein |
| CN109507174A (en) * | 2019-01-16 | 2019-03-22 | 济南大学 | Preparation based on the compound ZnO nanoparticle quenching Particles in Electrochemiluminescofce ofce Luminol sensor of curcumin |
Non-Patent Citations (2)
| Title |
|---|
| Ferritin-Based Electrochemiluminescence Nanosurface Energy Transfer System for Procalcitonin Detection Using HWRGWVC Heptapeptide for Site-Oriented Antibody Immobilization;Lei Yang et al;《American Chemical Society》;20190510;第7145-7152页 * |
| Macroporous graphene capped Fe3O4 for amplified electrochemiluminescence immunosensing of carcinoembryonic antigen detection based on CeO2@TiO2;Lei Yang等;《Biosensors and Bioelectronic》;20171231;第1-25页 * |
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