CN110779976B - Cancer blank antigen detection method based on UiO-66(NH2) -silver sulfide composite material - Google Patents
Cancer blank antigen detection method based on UiO-66(NH2) -silver sulfide composite material Download PDFInfo
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Abstract
The invention relates to the field of biological detection, in particular to a cancer blank antigen detection method based on a UiO-66(NH2) -silver sulfide composite material, which comprises the following steps of S1: synthesis of UiO-66(NH 2); s2: modifying a layer of UiO-66(NH2) on the FTO glass; s3: growing Ag2S on the surface of the UiO-66(NH2) compound to obtain a UiO-66(NH2) -silver sulfide composite material; s4, installing the FTO glass loaded with the UiO-66(NH2) -silver sulfide composite material on an electrode, placing the electrode in a PBS buffer solution of ascorbic acid, electrifying, and carrying out electrochemical reaction to detect the level of the cancer embryo antigen in the ascorbic acid. The detection method has the advantages of low detection cost, good reliability of the detection result, lower detection limit and wider linear range.
Description
Technical Field
The invention relates to the field of biological detection, in particular to a cancer blank antigen detection method based on a UiO-66(NH2) -silver sulfide composite material.
Background
CEA is one of the embryonic antigens, a normal component, as it is produced by embryonic tissue at the embryonic development stage. As the embryo grows, the content of embryo antigens in the later period of the embryo is reduced, and the antigens gradually disappear or only exist in a trace amount after birth. When cells become cancerous, the antigen can be synthesized again, expressed on the surface of tumor cells and secreted into blood, and the content of the antigen is always in positive correlation with the degree of cell malignancy, so that CEA is often used as an important index for detecting tumors in clinical diagnosis. The existing immune methods for determining CEA mainly comprise enzyme-linked immunoassay, fluorescence immunoassay, radioimmunoassay and the like. These conventional immunoassay methods are generally complex to operate, expensive, and unsuitable for real-time detection.
Photoelectrochemical detection, a method of photoelectrochemical analysis that combines the advantages of Photoelectrochemical (PEC) bioanalytical sensitivity and specificity of immune molecules. In the prior art, the photoelectrochemistry detection of CEA has the following defects that on one hand, the photoelectric activity and the stability of the sensor are required to be improved, and the reliability of the detection result is not high; in the second aspect, the detection limit is high and the linear range is narrow.
Disclosure of Invention
In order to solve the problems, the invention provides a cancer blank antigen detection method based on the UiO-66(NH2) -silver sulfide composite material, which has the advantages of low detection cost, good detection result reliability, lower detection limit and wider linear range.
The technical scheme adopted by the invention is as follows: cancer blank antigen detection method based on UiO-66(NH2) -silver sulfide composite material, S1: synthesis of UiO-66(NH 2); s2: modifying a layer of UiO-66(NH2) on the FTO glass; s3: growing Ag2S on the surface of the UiO-66(NH2) compound to obtain a UiO-66(NH2) -silver sulfide composite material; s4, installing the FTO glass loaded with the UiO-66(NH2) -silver sulfide composite material on an electrode, placing the electrode in a PBS buffer solution of ascorbic acid, electrifying, and carrying out electrochemical reaction to detect the level of the cancer embryo antigen in the ascorbic acid.
The technical proposal is further improved in that in S1, UiO-66 (NH)2) The synthesis method comprises the following steps: a. ultrasonically dissolving zirconium tetrachloride and 2-amino terephthalic acid in a DMF solution by using a solvothermal reaction method, transferring the solution into a reaction kettle, and reacting for 12 to 36 hours at the temperature of between 10 and 200 ℃; b. centrifuging a light yellow product obtained after the reaction in the reaction kettle in the step a, washing the light yellow product for 1-5 times by using a washing solvent, drying the product in vacuum at 10-200 ℃, and standing the dried product to obtain UiO-66 (NH)2) And (3) solid powder.
The technical proposal is further improved in that in S2, UiO-66 (NH)2) The method for modifying the FTO glass comprises the following steps: a. cutting the FTO glass into sheets, ultrasonically cleaning for 1-5 min, b, drying the cleaned sheet FTO glass in the step a by using nitrogen, and taking 0.5mg mL-1～3.5mg mL-1UiO-66 (NH)2) Uniformly dropwise adding the mixture on the surface of FTO glass, and naturally drying at room temperature.
The technical proposal is further improved in that in S3, in UiO-66 (NH)2) Growing Ag on the surface of compound2The method of S is as follows: a. taking AgNO3Mixing the ethanol and water to prepare a cation precursor solution; b. mixing ethanol containing thiourea with water to obtain an anion precursor solution; c. modification of UiO-66 (NH) in S22) The FTO glass is sequentially immersed into the cation precursor solution and the anion precursor solution, the circulation is carried out for 3 to 9 times, the immersion time is 3 to 7min each time, so that the Ag2S nano-particles grow on UiO-66 (NH)2) A surface; d. cleaning and drying to obtain the UiO-66(NH2) -silver sulfide composite material.
The technical scheme is further improved in that in S1, the reaction temperature in the reaction kettle is 120 ℃, the reaction time is 24 hours, the washing solvent is methanol, the washing times are 3 times, and the vacuum drying temperature is 80 ℃.
The technical proposal is further improved in that in S2, the FTO glass is ultrasonically cleaned in absolute ethyl alcohol and deionized water for 3 minutes in sequence, and 2.5mg mL of the FTO glass is taken-1UiO-6 of6(NH2) Dropwise addition was carried out.
The technical proposal is further improved in that in S3, the cation precursor solution contains AgNO3The volume ratio of the ethanol to the water is 4:1, and the volume ratio of the ethanol containing thiourea in the anion precursor solution to the water is 4: 1; circulating for 7 times, and soaking for 5min each time; ag in UiO-66(NH2) -silver sulfide composite material2The quantum dots of S are 8 nm-12 nm.
The technical scheme is further improved in that in S4, a working electrode in the electrochemical reaction is FTO glass loaded with UiO-66(NH2) -silver sulfide composite material, a platinum sheet is used as a counter electrode, a saturated calomel electrode is used as a reference electrode, a detection light source is a 500W xenon lamp, and the detection potential is 0.2V.
The invention has the beneficial effects that:
1. on one hand, the UiO-66(NH2) -silver sulfide composite material can utilize the semiconductor material with a narrow forbidden band to have better absorbance in a visible light region and avoid the inactivation of biomolecules caused by ultraviolet light, so the composite material has good chemical stability, lower detection limit, wider linear range and excellent physical and chemical properties. In the second aspect, UiO-66(NH2) -silver sulfide is used as a photoelectric sensor to detect the level of the cancer embryo antigen, and the method is simple, low in detection cost, high in detection sensitivity and high in detection precision, and is suitable for large-scale popularization. In a third aspect, the UiO-66 (NH) of the present invention is comparable to conventional wide bandgap semiconductor materials2) Silver sulfide composite, combined with UiO-66 (NH)2) And the light absorption frequency of silver sulfide, materials with different band gap widths are combined, so that a visible light region also has better absorbance and good photoelectrochemical stability, the composite material is applied to photoelectrochemical detection of a tumor marker carcinoembryonic antigen, and the result proves that the photoelectric sensor has lower detection limit and wider linear range.
2. In S1, UiO-66 (NH) was synthesized2) When the method is used, the methanol is adopted to clean a light yellow product in the reaction kettle, and the boiling point of the methanol is 64 ℃, so that the volatility is good, the methanol is ensured to be completely volatilized in the washing process, the purity of the product is improved, and the chemical stability of the product is further improved. After cleaning, vacuum was applied at 80 deg.CDrying to ensure that the UiO powder is completely dried, and further improving the UiO-66 (NH)2) The purity of the product is beneficial to improving the chemical stability of the finally obtained UiO-66(NH2) -silver sulfide, and the stability and reliability of the detection result are ensured.
3. A layer of UiO-66 (NH) is modified on the FTO glass2) In the process, the FTO glass is ultrasonically cleaned in absolute ethyl alcohol and deionized water for 3 minutes in sequence, and 2.5mg mL of the FTO glass is taken-1UiO-66 (NH)2) Dropwise adding; ensuring the cleanness of the conductive surface of the glass, avoiding the contamination of foreign matters and preventing the foreign matters from blocking the active substance UiO-66 (NH)2) The load on the FTO improves the purity of the finally obtained UiO-66(NH2) -silver sulfide, ensures the stability and reliability of the chemical property of the silver sulfide, and ensures the reliability of the detection result.
4. In S3, the selected anion precursor solution and cation precursor solution have low material cost, no pollution and suitability for batch synthesis, and meanwhile, the anion precursor solution and the cation precursor solution sequentially carry UiO-66 (NH)2) The FTO glass is soaked to ensure Ag2S nano-particles can be completely grown on UiO-66 (NH)2) On the surface, the prepared UiO-66(NH2) -silver sulfide has high purity, ensures the stability and reliability of the chemical property and the reliability of the detection result.
5. The quantum dot of Ag2S in the UiO-66(NH2) -silver sulfide composite material is 8 nm-12 nm, preferably 10nm, the material band gap width of the quantum dot is small, about 1eV, the quantum dot is easy to absorb light and excite to generate photoelectrons, so that the photoelectric reaction is better carried out to obtain the CEA level, and the stability and the reliability of the detection result are further ensured.
6. In the detection process of the CEA level, CEA solutions with different concentrations are dripped on a prepared FTO electrode, a period of incubation is carried out, then photoelectric response is measured in an AA-containing PBS solution, because CEA has a certain shading effect, the electron donor in the solution is hindered from improving the electron path to the photoelectric material, so that the photo-generated electron hole recombination is caused, the photocurrent is reduced, namely the higher the concentration of CEA is, the lower the photoelectric response is. The higher the energy output by the detection light source, the larger the obtained photocurrent, and when the detection light source is a 500W xenon lamp and the detection potential is 0.2V, the photocurrent is strongest, the sensitivity to CEA is highest, and the detection precision is high.
Drawings
FIG. 1 is an SEM image of the prepared UiO-66(NH2) -silver sulfide;
FIG. 2 is a graph of the photoelectrochemical stability test of UiO-66(NH2) -silver sulfide/FTO in 0.1M PBS containing 0.1M ascorbic acid.
Detailed Description
The invention will be further described with reference to the accompanying drawings.
Example 1: synthesis of UiO-66(NH2) -silver sulfide composite material
S1: synthesizing UiO-66(NH2), specifically, a, ultrasonically dissolving zirconium tetrachloride and 2-amino terephthalic acid in a DMF solution by using a solvothermal reaction method, transferring the solution to a reaction kettle, and reacting for 24 hours at 120 ℃; b. centrifuging a light yellow product obtained after the reaction in the reaction kettle in the step a, washing the light yellow product for 3 times by using a washing solvent, drying the light yellow product in vacuum at 80 ℃, and standing the dried product to obtain UiO-66 (NH)2) And (3) solid powder.
S2: modifying a layer of UiO-66(NH2) on FTO glass, specifically, a, cutting the FTO glass into sheets, ultrasonically cleaning the sheets in absolute ethyl alcohol and deionized water for 3 minutes in sequence, b, drying the cleaned sheets of the FTO glass in the step a by using nitrogen, and taking 2.5mg mL of the dried sheets of the FTO glass-1UiO-66 (NH)2) Uniformly dropwise adding the mixture on the surface of FTO glass, and naturally drying at room temperature.
S3: ag2S grows on the surface of the UiO-66(NH2) compound to obtain the UiO-66(NH2) -silver sulfide composite material. The method specifically comprises the following steps: a. taking AgNO3Mixing the ethanol and water to prepare a cation precursor solution; b. mixing ethanol containing thiourea with water to obtain an anion precursor solution; c. modification of UiO-66 (NH) in S22) Sequentially immersing the FTO glass into a cation precursor solution and an anion precursor solution, circulating for 7 times, and soaking for 5min each time to ensure that the Ag is2S nano-particles grow on UiO-66 (NH)2) A surface; d. cleaning and blow-drying to obtain the UiO-66(NH2) -silver sulfide composite material. In S3, the cation precursor solution contains AgNO3The volume ratio of the ethanol to the water is 4:1, and the volume ratio of the ethanol containing thiourea in the anion precursor solution to the water is 4: 1; circulating for 7 times, and soaking for 5min each time; UiO-66(NH2) -sulfurizationAg in silver composite material2The quantum dots of S are 10 nm.
The SEM image of the prepared UiO-66(NH2) -silver sulfide was tested, and the results are shown in FIG. 1. As can be seen from fig. 1, the UiO-66(NH2) -silver sulfide synthesized in this embodiment has a narrow bandgap, and a semiconductor material with a narrow bandgap can have a good absorbance in the visible light region to avoid the deactivation of biomolecules caused by ultraviolet light.
Example 2: CEA determination using UiO-66(NH2) -silver sulfide composite
The method comprises the steps of taking UiO-66(NH2) -silver sulfide in example 1, mounting FTO glass loaded with UiO-66(NH2) -silver sulfide composite material on an electrode, placing the electrode in a PBS buffer solution of ascorbic acid, electrifying, and carrying out electrochemical reaction to detect the level of cancer embryo antigens in the ascorbic acid.
During detection, a working electrode is FTO glass loaded with UiO-66(NH2) -silver sulfide composite material during electrochemical reaction, a platinum sheet is used as a counter electrode, a saturated calomel electrode is used as a reference electrode, a detection light source is a 500W xenon lamp, and the detection potential is 0.2V. The results of the detection are shown in FIG. 2.
As can be seen from FIG. 2, UiO-66 (NH) with a narrow bandgap compared to conventional wide bandgap semiconductor materials2) Silver sulfide composite, combined with UiO-66 (NH)2) And the light absorption frequency of silver sulfide, and materials with different band gap widths are combined, so that the visible light region also has better absorbance and good photoelectrochemical stability. The result of applying the composite material to the photoelectrochemical detection of the tumor marker carcinoembryonic antigen proves that the photoelectric sensor has lower detection limit and wider linear range.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (6)
1. The cancer blank antigen detection method based on the UiO-66(NH2) -silver sulfide composite material is characterized by comprising the following steps: comprising the steps of S1: synthesis of UiO-66(NH 2); s2: modifying a layer of UiO-66(NH2) on the FTO glass; s3: growing Ag2S on the surface of the UiO-66(NH2) compound to obtain a UiO-66(NH2) -silver sulfide composite material; s4, installing the FTO glass loaded with the UiO-66(NH2) -silver sulfide composite material on an electrode, placing the electrode in a PBS buffer solution of ascorbic acid, electrifying, and carrying out electrochemical reaction to detect the level of cancer embryo antigens in the ascorbic acid; in the S2, UiO-66 (NH)2) The method for modifying the FTO glass comprises the following steps: a. cutting the FTO glass into sheets, ultrasonically cleaning for 1-5 min, b, drying the cleaned sheet FTO glass in the step a by using nitrogen, and taking 0.5mg mL-1～3.5mg mL-1UiO-66 (NH)2) Uniformly dropwise adding the mixture on the surface of FTO glass, and naturally drying at room temperature; in said S3, in UiO-66 (NH)2) Growing Ag on the surface of compound2The method of S is as follows: a. taking AgNO3Mixing the ethanol and water to prepare a cation precursor solution; b. mixing ethanol containing thiourea with water to obtain an anion precursor solution; c. modification of UiO-66 (NH) in S22) The FTO glass is sequentially immersed into the cation precursor solution and the anion precursor solution, the circulation is carried out for 3 to 9 times, the immersion time is 3 to 7min each time, so that the Ag2S nanoparticles are grown on UiO-66 (NH)2) A surface; d. cleaning and blow-drying to obtain UiO-66(NH2) -silver sulfide composite material; the cancer embryo antigen detection method based on the UiO-66(NH2) -silver sulfide composite material is not used for diagnosis and treatment of diseases.
2. The method for detecting the carcinoembryonic antigen based on the UiO-66(NH2) -silver sulfide composite material of claim 1, wherein the method comprises the following steps: in S1, UiO-66 (NH)2) The synthesis method comprises the following steps: a. ultrasonically dissolving zirconium tetrachloride and 2-amino terephthalic acid in a DMF solution by using a solvothermal reaction method, transferring the solution into a reaction kettle, and reacting for 12 to 36 hours at the temperature of between 10 and 200 ℃; b. centrifuging a light yellow product obtained after the reaction in the reaction kettle in the step a, washing the light yellow product for 1-5 times by using a washing solvent, drying the product in vacuum at 10-200 ℃, and standing the dried product to obtain UiO-66 (NH)2) And (3) solid powder.
3. The method for detecting the carcinoembryonic antigen based on the UiO-66(NH2) -silver sulfide composite material of claim 2, wherein: in S1, the reaction temperature in the reaction kettle is 120 ℃, the reaction time is 24h, the washing solvent is methanol, the washing times are 3 times, and the vacuum drying temperature is 80 ℃.
4. The method for detecting the carcinoembryonic antigen based on the UiO-66(NH2) -silver sulfide composite material of claim 3, wherein: in S2, the FTO glass is ultrasonically cleaned in absolute ethyl alcohol and deionized water for 3 minutes in sequence, and 2.5mg mL of the FTO glass is taken-1UiO-66 (NH)2) Dropwise addition was carried out.
5. The method for detecting the carcinoembryonic antigen based on the UiO-66(NH2) -silver sulfide composite material of claim 4, wherein: in S3, the cation precursor solution contains AgNO3The volume ratio of the ethanol to the water is 4:1, and the volume ratio of the ethanol containing thiourea in the anion precursor solution to the water is 4: 1; circulating for 7 times, and soaking for 5min each time; ag in UiO-66(NH2) -silver sulfide composite material2The quantum dots of S are 8 nm-12 nm.
6. The method for detecting carcinoembryonic antigen based on UiO-66(NH2) -silver sulfide composite material according to claim 5, wherein: in S4, a working electrode is FTO glass loaded with UiO-66(NH2) -silver sulfide composite material during electrochemical reaction, a platinum sheet is used as a counter electrode, a saturated calomel electrode is used as a reference electrode, a detection light source is a 500W xenon lamp, and the detection potential is 0.2V.
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| CN111307902B (en) * | 2020-03-12 | 2024-01-12 | 东莞理工学院 | Based on Fe 2 O 3 Carcinoembryonic antigen detection method of material monolayer photoelectrochemical sensor |
| CN113791127B (en) * | 2021-09-26 | 2022-09-02 | 东莞理工学院 | Nano-gold-loaded tungsten oxide-silver sulfide composite material and preparation method and application thereof |
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