CN114674891A - Construction of hollow structures in combination with electronic consumption strategy sensors - Google Patents

Construction of hollow structures in combination with electronic consumption strategy sensors Download PDF

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CN114674891A
CN114674891A CN202210268542.1A CN202210268542A CN114674891A CN 114674891 A CN114674891 A CN 114674891A CN 202210268542 A CN202210268542 A CN 202210268542A CN 114674891 A CN114674891 A CN 114674891A
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颜梅
苗培
张晶
秦成坤
马廷滨
刘明霞
吕艳锋
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University of Jinan
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Abstract

The invention discloses a method for constructing a hollow structure combined with an electron consumption strategy sensor, which firstly synthesizes hollow structure ZnCdS/ZnIn2S4A heterojunction, which can facilitate light collection by multiple light scattering/reflection, while synergy between the hollow structure and the intimate contact heterojunction interface can facilitate visible light harvesting, accelerate charge transfer, and inhibit recombination of photogenerated carriers; in addition, AuNPs-AAO is modified on the secondary antibody to be used as a signal amplification carrier, the quenching effect induced by AAO is used for inhibiting photocurrent signals, and under the action, AAO catalysis is carried outOxidation of AA results in a reduction in AA usage, thereby greatly reducing photocurrent response.

Description

Construction of hollow structures in combination with electronic consumption strategy sensors
Technical Field
The invention relates to the field of quantitative detection of prostate specific antigen, in particular to construction of a hollow structure and electron consumption strategy combined sensor.
Background
Prostate cancer is a malignant epithelial tumor of the prostate gland and is common in men worldwide. Prostate lesions are asymptomatic in the early stages and can be fatal when severe. Therefore, the early detection and treatment of the prostatic cancer have very important significance for diagnosing malignant tumors, a new simple, quick, sensitive and high-selectivity method for detecting the malignant tumor biomarkers is established, and the method has very important values for early detection and treatment effect evaluation of the malignant tumors.
Photoelectrochemical (PEC) immunosensors have received much attention because of their low background signal, high sensitivity, simple operation, inexpensive instrumentation, and the like. The method is a sensitive detection technology combining photoelectrochemistry and immunology, and has strong specificity of immunology and high sensitivity of photoelectrochemistry. Briefly, the detection mechanism of the PEC immunosensor is to convert the content of the substance to be detected into an optoelectronic signal by signal amplification under illumination conditions.
Disclosure of Invention
The object of the present invention is to construct a photoelectrochemical sensor for prostate specific antigen detection using a hollow structure in combination with an electron depletion strategy sensor.
In order to solve the technical problem, the invention is realized by the following measures: the construction of the hollow structure combined with an electronic consumption strategy sensor is characterized by comprising the following steps:
(1) synthesis of ZIF-8: 2.36 g of Zn (NO)3)2·6H2O is dissolved in 200 mL of methanol, 1.32 g of 2-methylimidazole is rapidly added to the solution under stirring, and then aging is carried out at room temperature for 11 hours; finally, it was washed 3 times with ethanol and dried under vacuum at 70 ℃ for 12 h.
(2) Synthesis of ZIF-8 derived hollow ZnCdS: mixing 50 mL of ethanol solution with 20 mg of ZIF-8 synthesized in the step (1) and 400 mg of thioacetamide, transferring the mixture into a 100 mL autoclave, keeping the mixture at 150 ℃ for 1 hour, centrifuging the mixture, collecting a product, and washing the product with ethanol to obtain a purple product; thereafter 20 mg of violet product and 78 mg of CdCl 2Dissolved in 25 mL of ethanol and magnetically stirred, the mixture was transferred to a 50 mL autoclave, heated at 160 ℃ for 4 h, and the resulting yellow precipitate was collected by centrifugation, washed with ethanol and then dried under vacuum at 70 ℃ for 12 h.
(3) Synthesis of hollow ZnCdS/ZnIn2S4And (3) heterojunction: weighing 0.2 g of the powder obtained in the step (2), dispersing in 20 mL of ethanol, and carrying out ultrasonic treatment for 30 min; then, 1 mmol of ZnCl2、2 mmol InCl3·4H2O and 8 mmol thioacetamide are added into the suspension and stirred intensively for 30 min; sealing the obtained mixture in a 50 mL autoclave lined with polytetrafluoroethylene, and keeping the temperature at 120 ℃ for 2 h; the product is extracted by centrifugation, washed with deionized water and ethanol, and dried at 60 ℃.
(4) Synthesis of ITO/ZnCdS/ZnIn2S4An electrode: the conductive glass is Indium Tin Oxide (ITO), is cut into strips of 4.0 multiplied by 0.5 cm, is ultrasonically cleaned for 5 min by acetone solution, secondary distilled water and absolute ethyl alcohol in sequence, and is dried under nitrogen for standby; the concentration of ZnCdS/ZnIn synthesized in the step (3) is 2.0 mg/mL2S4Dripping the mixture on ITO glass, and drying the mixture at 60 ℃ to obtain ITO/ZnCdS/ZnIn2S4And an electrode.
(5) Synthesis of AuNPs: under continuous stirring, 1.5 mL of 25 mmol/L HAuCl was added 4And 5 mL of sodium citrate with a concentration of 10 mmol/L are mixed in a round-bottom flask; thereafter, 1.5 mL of 0.1 mol/L NaBH4Quickly injected into the above solution, then it turns orange-red, indicating particle formation; after that, the mixed solution was stirred at room temperature for another 6 hours, and the color changed from orange red to wine red; finally, the resulting AuNPs solution was stored at 4 ℃.
(6) Synthesizing AuNPs-AAO: first, 0.2 mol of K is added dropwise2CO3Stirring and adjusting the pH value of the Au NPs solution synthesized in the step (5) to 8.2; subsequently, 40. mu.L of 0.8 mg/mL Ascorbic Acid Oxidase (AAO) was added to 2.0 mL of the alkaline Au NPs solution, respectively, incubated for 2 hours, and the solution mixture was centrifuged at 10000 rpm for 20 min and washed several times to remove residues.
(7) Synthesis of Ab 2-AuNPs-AAO: adding 1 mL of secondary antibody, namely Ab2, with the concentration of 10 mu g/mL into the synthesized product in the step (6), incubating at 4 ℃ for 2 h, and washing with phosphate buffer solution with the pH of 7.4 for 3 times to remove the Ab2 without complexing, thereby obtaining Ab 2-AuNPs-AAO.
(8) Construction of a photoelectric sensor (PEC): rinsing ITO/ZnCdS/ZnIn with ultrapure water2S4Electrodes, 6 μ L primary antibody, Ab1, at a concentration of 10 μ g/mL was then incubated at 4 ℃ for 16 h, rinsed thoroughly 3 times with phosphate buffer pH 7.4; continuously dripping 20 mu L of 3% bovine serum albumin to block the non-specific binding sites, thoroughly washing the non-specific binding sites for 3 times by using a phosphate buffer solution with pH 7.4, dripping 20 mu L of prostate antigens with different concentrations to the surface of the electrode, incubating the electrode for 30 min at room temperature, and washing the electrode for 3 times by using a phosphate buffer solution with pH 7.4; 20 μ L of Ab2-AuNPs-AAO synthesized in step (7) was further added dropwise and incubated at room temperature for 4 h.
(9) Electrochemical detection of the photoelectric sensor: and (3) taking the modified electrode processed in the step (7) as a working electrode, taking the counter electrode as a platinum wire electrode, taking the reference electrode as an Ag/AgCl electrode, taking the bias voltage value as 0V, taking a xenon lamp as a light source for stimulation, taking an electrolytic cell as a phosphate buffer solution system (containing 1 mol/L of ascorbic acid) with the pH value of 7.4, and measuring a current I-T curve to detect the photoelectric property.
The invention has the beneficial effects that:
(1) the method has the advantages of low cost, simple experimental operation and easy control of reaction conditions.
（2）ZnCdS/ZnIn2S4The heterojunction hierarchical hollow structure can facilitate light collection by multiple light scattering/reflection.
（3）ZnCdS/ZnIn2S4The synergy between the heterojunction hollow structure and the intimate contact heterojunction interface may facilitate visible light harvesting, accelerate charge transport, and inhibit recombination of photogenerated carriers.
(4) The PEC signal is suppressed by AAO-induced quenching effects, which act to reduce the amount of AA used by catalytic oxidation of AA, thereby greatly reducing the photocurrent response.
Detailed Description
In order to further understand the invention, the technical scheme is implemented by combining the embodiment, and the specific implementation mode is given as follows:
(1) synthesizing ZIF-8: 2.36 g of Zn (NO)3)2·6H2O is dissolved in 200 mL of methanol, 1.32 g of 2-methylimidazole is rapidly added to the solution under stirring, and then aging is carried out at room temperature for 11 hours; finally, it was washed 3 times with ethanol and dried under vacuum at 70 ℃ for 12 h.
(2) Synthesis of ZIF-8 derived hollow ZnCdS: mixing 50 mL of ethanol solution with 20 mg of ZIF-8 synthesized in the step (1) and 400 mg of thioacetamide, transferring the mixture into a 100 mL autoclave, keeping the mixture at 150 ℃ for 1 hour, centrifuging the mixture, collecting a product, and washing the product with ethanol to obtain a purple product; thereafter 20 mg of the violet product and 78 mg of CdCl2Dissolved in 25 mL of ethanol, after magnetic stirring, the mixture was transferred to a 50 mL autoclave, heated at 160 ℃ for 4 h, and the resulting yellow precipitate was collected by centrifugation, washed with ethanol and then dried under vacuum at 70 ℃ for 12 h.
(3) Synthesis of hollow ZnCdS/ZnIn2S4Heterojunction: weighing 0.2 g of the powder obtained in the step (2), dispersing in 20 mL of ethanol, and carrying out ultrasonic treatment for 30 min; then, 1 mmol of ZnCl2、2 mmol InCl3·4H2O and 8 mmol thioacetamide are added into the suspension and stirred intensively for 30 min; sealing the obtained mixture in a 50 mL autoclave lined with polytetrafluoroethylene, and keeping the temperature at 120 ℃ for 2 h; the product is extracted by centrifugation, washed with deionized water and ethanol, and dried at 60 ℃.
(4) Synthesis of ITO/ZnCdS/ZnIn2S4An electrode: the conductive glass is Indium Tin Oxide (ITO), is cut into strips of 4.0 multiplied by 0.5 cm, is ultrasonically cleaned for 5 min by acetone solution, secondary distilled water and absolute ethyl alcohol in sequence, and is dried under nitrogen for standby; the concentration of ZnCdS/ZnIn synthesized in the step (3) is 2.0 mg/mL 2S4Dripping the mixture on ITO glass, and drying the mixture at the temperature of 60 ℃ to obtain ITO/ZnCdS/ZnIn2S4And an electrode.
(5) Synthesizing AuNPs: 1.5 mL of 25 mmol/L HAuCl were added with continuous stirring4And 5 mL ofMixing sodium citrate with the concentration of 10 mmol/L in a round-bottom flask; thereafter, 1.5 mL of 0.1 mol/L NaBH4Quickly injected into the above solution, then it turns orange-red, indicating particle formation; after that, the mixed solution was stirred at room temperature for another 6 hours, and the color changed from orange red to wine red; finally, the resulting AuNPs solution was stored at 4 ℃.
(6) Synthesizing AuNPs-AAO: first, 0.2 mol of K is added dropwise2CO3Stirring and adjusting the pH value of the Au NPs solution synthesized in the step (5) to 8.2; subsequently, 40. mu.L of 0.8 mg/mL Ascorbic Acid Oxidase (AAO) was added to 2.0 mL of the alkaline Au NPs solution, respectively, incubated for 2 hours, and the solution mixture was centrifuged at 10000 rpm for 20 min and washed several times to remove residues.
(7) Synthesis of Ab 2-AuNPs-AAO: adding 1 mL of secondary antibody, namely Ab2, with the concentration of 10 mu g/mL into the synthesized product in the step (6), incubating at 4 ℃ for 2 h, and washing with phosphate buffer solution with the pH of 7.4 for 3 times to remove the Ab2 without complexing, thereby obtaining Ab 2-AuNPs-AAO.
(8) Construction of a photoelectric sensor (PEC): rinsing ITO/ZnCdS/ZnIn with ultrapure water 2S4Electrodes, followed by 6 μ L primary antibody, Ab1, at a concentration of 10 μ g/mL, incubated at 4 ℃ for 16 h, rinsed thoroughly 3 times with phosphate buffer pH 7.4; continuously dripping 20 mu L of 3% bovine serum albumin to block the non-specific binding sites, thoroughly washing the non-specific binding sites for 3 times by using a phosphate buffer solution with pH 7.4, dripping 20 mu L of prostate antigens with different concentrations onto the surface of the electrode, incubating the electrode at room temperature for 30 min, and washing the electrode for 3 times by using a phosphate buffer solution with pH 7.4; mu.L of Ab2-AuNPs-AAO synthesized in step (7) was further added dropwise and incubated at room temperature for 4 hours.
(9) Electrochemical detection of the photoelectric sensor: the modified electrode treated in the step (7) is used as a working electrode, the counter electrode is a platinum wire electrode, the reference electrode is an Ag/AgCl electrode, the bias voltage value is 0V, a xenon lamp is used as a light source for stimulation, the electrolytic cell is a phosphate buffer solution system (containing 1 mol/L ascorbic acid) with the pH value of 7.4, the photoelectric property is detected by measuring a current I-T curve, the linear equation obtained is I = -1.04log (c) -10.44, the correlation coefficient is 0.992, and the detection limit is 0.04 pg/mL, so that the prostate specific antigen is detected with high sensitivity.

Claims (10)

1. The construction of the hollow structure combined with an electronic consumption strategy sensor is characterized by comprising the following steps:
(1) Synthesizing ZIF-8;
(2) synthesizing ZIF-8 derived hollow ZnCdS;
(3) synthesis of hollow ZnCdS/ZnIn2S4A heterojunction;
(4) construction of ITO/ZnCdS/ZnIn2S4An electrode;
(5) synthesizing AuNPs;
(6) synthesizing AuNPs-AAO;
(7) ab2-AuNPs-AAO is synthesized;
(8) construction of a photoelectric sensor (PEC);
(9) electrochemical detection of the photosensor.
2. The hollow structure of claim 1, in combination with the construction of an electronic consumption strategy sensor, is a synthetic ZIF-8, characterized by: 2.36 g of Zn (NO)3)2·6H2O is dissolved in 200 mL of methanol, 1.32 g of 2-methylimidazole is rapidly added to the solution under stirring, and then aging is carried out at room temperature for 11 hours; finally, it was washed 3 times with ethanol and dried under vacuum at 70 ℃ for 12 h.
3. The synthesis of ZIF-8 derived hollow ZnCdS, according to the hollow structure of claim 1, in combination with the construction of electron-consuming strategic sensors, characterized by: mixing 50 mL of ethanol solution with 20 mg of ZIF-8 synthesized in the step (1) and 400 mg of thioacetamide, transferring the mixture into a 100 mL autoclave, keeping the mixture at 150 ℃ for 1 hour, centrifuging the mixture, collecting a product, and washing the product with ethanol to obtain a purple product; thereafter 20 mg of violet product and 78 mg of CdCl2Dissolved in 25 mL of ethanol, after magnetic stirring, the mixture was transferred to a 50 mL autoclave, heated at 160 ℃ for 4 h, and the resulting yellow precipitate was collected by centrifugation, washed with ethanol and then dried under vacuum at 70 ℃ for 12 h.
4. Hollow structure according to claim 1 combined with the construction of an electron consumption strategy sensor, synthesizing a hollow ZnCdS/ZnIn2S4A heterojunction, characterized by: weighing 0.2 g of the powder obtained in the step (2), dispersing in 20 mL of ethanol, and carrying out ultrasonic treatment for 30 min; then, 1 mmol of ZnCl was added2、2 mmol InCl3·4H2O and 8 mmol thioacetamide are added into the suspension and stirred vigorously for 30 min; sealing the obtained mixture in a 50 mL autoclave lined with polytetrafluoroethylene, and keeping the temperature at 120 ℃ for 2 h; the product is extracted by centrifugation, washed with deionized water and ethanol, and dried at 60 ℃.
5. Hollow structure according to claim 1 combined with the construction of an electron consumption strategy sensor, synthesizing ITO/ZnCdS/ZnIn2S4An electrode, characterized by: the conductive glass is Indium Tin Oxide (ITO), is cut into strips of 4.0 multiplied by 0.5 cm, is ultrasonically cleaned for 5 min by acetone solution, secondary distilled water and absolute ethyl alcohol in sequence, and is dried under nitrogen for standby; ZnCdS/ZnIn with the concentration of 2.0 mg/mL and synthesized in the step (3)2S4Dripping the mixture on ITO glass, and drying the mixture at 60 ℃ to obtain ITO/ZnCdS/ZnIn2S4And an electrode.
6. The hollow structure of claim 1 in combination with the construction of an electronic consumption strategy sensor, synthesized AuNPs, characterized by: 1.5 mL of 25 mmol/L HAuCl were added with continuous stirring 4And 5 mL of sodium citrate with the concentration of 10 mmol/L are mixed in a round-bottom flask; thereafter, 1.5 mL of 0.1 mol/L NaBH4Quickly injected into the above solution, then it turns orange-red, indicating particle formation; after that, the mixed solution was stirred at room temperature for another 6 hours, and the color changed from orange red to wine red; finally, the resulting AuNPs solution was stored at 4 ℃.
7. The hollow structure of claim 1, in combination with the construction of an electronic consumption strategy sensor, synthesized AuNPs-AAO, wherein: first, 0.2 mol of K is added dropwise2CO3Stirring and adjusting the pH value of the Au NPs solution synthesized in the step (5) to 8.2; subsequently, 40. mu.L of 0.8 mg/mL Ascorbic Acid Oxidase (AAO) was added to 2.0 mL of the alkaline Au NPs solution, respectively, incubated for 2 hours, and the solution mixture was centrifuged at 10000 rpm for 20 min and washed several times to remove residues.
8. The construction of the hollow structure-bound electron consumption strategy sensor of claim 1, synthesizing Ab2-AuNPs-AAO, characterized by: adding 1 mL of secondary antibody, namely Ab2, with the concentration of 10 mu g/mL into the synthesized product in the step (6), incubating at 4 ℃ for 2 h, and washing with phosphate buffer solution with the pH of 7.4 for 3 times to remove the Ab2 without complexing, thereby obtaining Ab 2-AuNPs-AAO.
9. The hollow structure according to claim 1, in combination with the construction of an electronic consumption strategy sensor, a photoelectric sensor (PEC), characterized in that: rinsing ITO/ZnCdS/ZnIn with ultrapure water2S4Electrodes, 6 μ L primary antibody, Ab1, at a concentration of 10 μ g/mL was then incubated at 4 ℃ for 16 h, rinsed thoroughly 3 times with phosphate buffer pH 7.4; continuously dripping 20 mu L of 3% bovine serum albumin to block the non-specific binding sites, thoroughly washing the non-specific binding sites for 3 times by using a phosphate buffer solution with pH 7.4, dripping 20 mu L of prostate antigens with different concentrations to the surface of the electrode, incubating the electrode for 30 min at room temperature, and washing the electrode for 3 times by using a phosphate buffer solution with pH 7.4; 20 μ L of Ab2-AuNPs-AAO synthesized in step (7) was further added dropwise and incubated at room temperature for 4 h.
10. The hollow structure of claim 1 in combination with the construction of an electronic depletion strategy sensor, electrochemical detection by a photoelectric sensor, characterized by: and (3) taking the modified electrode processed in the step (7) as a working electrode, taking the counter electrode as a platinum wire electrode, taking the reference electrode as an Ag/AgCl electrode, taking the bias voltage value as 0V, taking a xenon lamp as a light source for stimulation, taking an electrolytic cell as a phosphate buffer solution system (containing 1 mol/L of ascorbic acid) with the pH value of 7.4, and measuring a current I-T curve to detect the photoelectric property.
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CN103262210A (en) * 2010-09-10 2013-08-21 维尔雷思科技有限公司 Methods of fabricating optoelectronic devices using layers detached from semiconductor donors and devices made thereby
CN110787814A (en) * 2019-11-07 2020-02-14 汕头大学 Layered hollow ZnCdS/MoS2Heterojunction cage and preparation and application thereof
CN113176314A (en) * 2021-03-09 2021-07-27 济南大学 Based on g-C3N4/Mo:BiVO4And CuS device preparation

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Title
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