CN114674894A - Construction of GQDs @ ZIF-8 as signal quencher sensor - Google Patents

Construction of GQDs @ ZIF-8 as signal quencher sensor Download PDF

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CN114674894A
CN114674894A CN202210268650.9A CN202210268650A CN114674894A CN 114674894 A CN114674894 A CN 114674894A CN 202210268650 A CN202210268650 A CN 202210268650A CN 114674894 A CN114674894 A CN 114674894A
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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 construction method of a sensor based on GQDs @ ZIF-8 as a signal quencher, which firstly synthesizes Bi4NbO8Cl/SnIn4S8Hetero-junction of which Bi4NbO8Cl as a silicon-based perovskite has visible light absorption properties, good stability and water resistance, and [ Bi ]2O2]And [ NbO ]4]The layered structure of the sequence is such that Bi4NbO8The carriers of Cl can be rapidly transferred; constructing Bi because of the appropriate energy band position4NbO8Cl/SnIn4S8The heterojunction can effectively improve the charge separation efficiency. In addition, the GQDs @ ZIF-8 polyhedron modified on the secondary antibody is used as a signal quencher, so that ITO/Bi can be inhibited through a steric hindrance effect4NbO8Cl/SnIn4S8The photocurrent signal of the electrode can also be used as a mimic of peroxidase to catalyze the precipitation reaction of 4-chloro-1-naphthol, so that the photocurrent signal is obviously reduced to achieve the effect of improving the sensitivity of the sensor.

Description

Construction of GQDs @ ZIF-8 as signal quencher sensor
Technical Field
The invention relates to the field of quantitative detection of prostate specific antigen, in particular to construction of a sensor taking GQDs @ ZIF-8 as a signal quencher.
Background
Along with the continuous deterioration of the environment, the probability of cancer is more and more, and the existing modes of radiotherapy, chemotherapy and the like can also cause serious damage to normal cells while treating the cancer, so that the realization of early detection and treatment of the cancer plays an important role in diagnosing malignant tumors, a new simple, quick, sensitive and good-selectivity method for detecting malignant tumor biomarkers is established, and the method has important values on early detection and treatment effect evaluation of the malignant tumors.
Photoelectrochemistry has attracted extensive research as a promising analytical method. Compared with traditional electrochemistry, PEC has the advantages of high sensitivity, good selectivity and low measurement cost due to the separation of the excitation light source and the output signal. Therefore, the method is widely applied in the fields of disease diagnosis, food safety detection, environmental protection and the like due to high sensitivity. The photoelectrochemical immunosensor outputs signals expressed by sensitive biological materials into electric signals by fixing the sensitive biological materials such as enzymes, antigens, antibodies, DNA and other active substances as recognition elements. The specific recognition of the biological material enables the photoelectrochemical immunosensor to have good specificity and sensitivity for the diagnosis of tumors.
Disclosure of Invention
The invention aims to construct a photoelectrochemical sensor for detecting prostate specific antigen by using GQDs @ ZIF-8 as a signal quencher sensor.
In order to solve the technical problem, the invention is realized by the following measures: the construction of the GQDs @ ZIF-8 as a signal quencher sensor is characterized by comprising the following steps:
(1) synthesis of Bi4NbO8Cl: simply mix 1.3 g of BiOCl, 0.66 g of Nb2O53.49 g of Bi2O, 2.92 g of NaCl and 3.7 g of KCl powder are ground for 10 min, and then the uniformly mixed powder is calcined at 700 ℃ for 3 h, washed with water at 90 ℃ for several times and then dried and collected.
(2) Synthesis of Bi4NbO8Cl/SnIn4S8And (3) heterojunction: 0.2 g of SnCl4·5H2O, 0.70 g of InCl3·4H2O and 0.45 g of thioacetamide in 70 mL of absolute ethanol; then 0.05 g of the powder synthesized in step (1) was dispersed in the above solution, heated to 70 ℃ by an oil bath and kept for 3 hours, and finally the resulting precipitate was centrifuged and washed 5 times with ethanol; finally, the product obtained was dried at 80 ℃ overnight to obtain Bi4NbO8Cl/SnIn4S8A heterojunction.
(3) Construction of ITO/Bi4NbO8Cl/SnIn4S8An 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; bi synthesized in the step (2) and having a concentration of 2.0 mg/mL4NbO8Cl/SnIn4S8Dripping on ITO glass, drying at 60 deg.C to obtain ITO/Bi4NbO8Cl/SnIn4S8And an electrode.
(4) Synthesis of GQDs: a mixture of 1.0 g glucose, 400. mu.L ethylenediamine and 200. mu.L HCl (37%, w/w) was added to 15 mL deionized water and stirred for 30 min; conveying the reaction mixture into a high-pressure kettle, and carrying out a hydrothermal process for 6 hours at the temperature of 200 ℃; the brown product from this step was centrifuged and dialyzed to remove large pieces of graphene oxide and the resulting GQDs solution was stored at 4 ℃.
(5) Synthesis of GQDs @ ZIF-8: 1.485 g of Zn (NO)3)2·6H2O and 3.28 g of 2-methylimidazole are dissolved in 50 mL of methanol respectively; 5 mL of the solution of step (4) was mixed with Zn (NO)3)2·6H2O solution is mixed with magnetic stirring for 0After 5 h, pouring into the mixture quickly; then stirring for 2 hours at room temperature; finally, the product was centrifuged, washed with methanol and dried overnight.
(6) Synthesis of SA-GQDs @ ZIF-8: adding 0.5 mL of 1 mg/mL Streptavidin (SA) to 2 mL of the product synthesized in step (5), stirring the mixture vigorously for 1 min and after 12 h of gentle shaking at 4 ℃, SA-labeled GQDs @ ZIF-8 are obtained; and stored at 4 ℃ protected from light for future use.
(7) Synthesis of Ab2-SA-GQDs @ ZIF-8: 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 for 2 h at 4 ℃, and washing for 3 times by using phosphate buffer solution with the pH of 7.4 to remove the Ab2 without complexing, thus obtaining Ab2-SA-GQDs @ ZIF-8.
(8) Construction of a photoelectric sensor (PEC): flushing ITO/Bi with ultrapure water4NbO8Cl/SnIn4S8Electrodes, 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; continuing to add 20. mu.L of the synthesized product of step (7), incubating at room temperature for 4H, and incubating the modified electrode in a solution containing 1 mM H 2O2Incubated with 10 mM 4 chloro-1-naphthol solution for 20 min.
(9) Electrochemical detection of the photoelectric sensor: and (3) taking the modified electrode processed in the step (8) 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 a phosphate buffer solution system (1 mol/L ascorbic acid) with the pH value of 7.4 as an electrolytic cell, and measuring a current I-T curve to detect the photoelectric property.
The invention has the beneficial effects that:
（1）Bi4NbO8cl, as a silicon-based perovskite, has visible light absorption performance and good stability and water resistance.
（2）[Bi2O2]And [ NbO ]4]The layered structure of the sequence is such that Bi4NbO8The carriers of Cl can migrate rapidly.
(3) Constructing Bi because of the appropriate energy band position4NbO8Cl/SnIn4S8The heterojunction can effectively improve the charge separation efficiency.
(4) GQDs @ ZIF-8 polyhedron is used as a signal quencher, and ITO/Bi can be inhibited through steric hindrance effect4NbO8Cl/SnIn4S8The photocurrent signal of the electrode can also be used as a mimic of peroxidase to catalyze the precipitation reaction of 4-chloro-1-naphthol, so that the photocurrent signal is obviously reduced to achieve the effect of improving the sensitivity of the sensor.
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:
(1) Synthesis of Bi4NbO8Cl: simply mix 1.3 g of BiOCl, 0.66 g of Nb2O53.49 g of Bi2O, 2.92 g of NaCl and 3.7 g of KCl powder are ground for 10 min, and then the uniformly mixed powder is calcined at 700 ℃ for 3 h, washed with water at 90 ℃ for several times and then dried and collected.
(2) Synthesis of Bi4NbO8Cl/SnIn4S8Heterojunction: 0.2 g of SnCl4·5H2O, 0.70 g of InCl3·4H2O and 0.45 g of thioacetamide in 70 mL of absolute ethanol; then 0.05 g of the powder synthesized in step (1) was dispersed in the above solution, heated to 70 ℃ by an oil bath and kept for 3 hours, and finally the resulting precipitate was centrifuged and washed 5 times with ethanol; finally, the product obtained was dried at 80 ℃ overnight to obtain Bi4NbO8Cl/SnIn4S8A heterojunction.
(3) Construction of ITO/Bi4NbO8Cl/SnIn4S8An electrode: the conductive glass is Indium Tin Oxide (ITO) glass, and is cut into 4.0Ultrasonic cleaning 0.5 cm strip with acetone solution, redistilled water and anhydrous alcohol for 5 min, and drying under nitrogen; bi synthesized in the step (2) and having a concentration of 2.0 mg/mL4NbO8Cl/SnIn4S8Dripping on ITO glass, drying at 60 deg.C to obtain ITO/Bi4NbO8Cl/SnIn4S8And an electrode.
(4) Synthesis of GQDs: a mixture of 1.0 g glucose, 400. mu.L ethylenediamine and 200. mu.L HCl (37%, w/w) was added to 15 mL deionized water and stirred for 30 min; conveying the reaction mixture into a high-pressure kettle, and carrying out a hydrothermal process for 6 hours at the temperature of 200 ℃; the brown product from this step was centrifuged and dialyzed to remove large pieces of graphene oxide and the resulting GQDs solution was stored at 4 ℃.
(5) Synthesis of GQDs @ ZIF-8: 1.485 g of Zn (NO)3)2·6H2O and 3.28 g of 2-methylimidazole are dissolved in 50 mL of methanol respectively; 5 mL of the solution of step (4) was mixed with Zn (NO)3)2·6H2Mixing the O solution for 0.5 h under magnetic stirring, and quickly pouring into the mixture; then stirring for 2 hours at room temperature; finally, the product was centrifuged, washed with methanol and dried overnight.
(6) Synthesis of SA-GQDs @ ZIF-8: adding 0.5 mL of 1 mg/mL Streptavidin (SA) to 2 mL of the product synthesized in step (5), stirring the mixture vigorously for 1 min and after 12 h of gentle shaking at 4 ℃, SA-labeled GQDs @ ZIF-8 are obtained; and stored at 4 ℃ protected from light for future use.
(7) Synthesis of Ab2-SA-GQDs @ ZIF-8: 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 for 2 h at 4 ℃, and washing for 3 times by using phosphate buffer solution with the pH of 7.4 to remove the Ab2 without complexing, thus obtaining Ab2-SA-GQDs @ ZIF-8.
(8) Construction of a photoelectric sensor (PEC): flushing ITO/Bi with ultrapure water4NbO8Cl/SnIn4S8Electrodes, 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; continue dispensing 20Blocking the non-specific binding sites by using 3% of microliter bovine serum albumin, thoroughly washing the nonspecific binding sites for 3 times by using a phosphate buffer solution with pH of 7.4, dropwise adding 20 microliter of prostate antigens with different concentrations to the surface of the electrode, incubating the prostate antigens at room temperature for 30 min, and washing the prostate antigens for 3 times by using a phosphate buffer solution with pH of 7.4; continuing to add 20. mu.L of the synthesized product of step (7), incubating at room temperature for 4H, and incubating the modified electrode in a solution containing 1 mM H 2O2Incubated with 10 mM 4 chloro-1-naphthol solution for 20 min.
(9) Electrochemical detection of the photoelectric sensor: the modified electrode treated in the step (8) 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, an electrolytic cell is a phosphate buffer solution system (ascorbic acid of 1 mol/L) of pH 7.4, the photoelectric property is detected by measuring a current I-T curve, the linear equation is obtained, I = -1.23log (c) -10.24, the correlation coefficient is 0.994, the detection limit is 0.07 pg/mL, and the prostate specific antigen is detected with high sensitivity.

Claims (10)

  1. The construction of GQDs @ ZIF-8 as a signal quencher sensor is characterized by comprising the following steps:
    (1) synthesis of Bi4NbO8Cl；
    (2) Synthesis of Bi4NbO8Cl/SnIn4S8A heterojunction;
    (3) construction of ITO/Bi4NbO8Cl/SnIn4S8An electrode;
    (4) synthesizing GQDs;
    (5) synthesizing GQDs @ ZIF-8;
    (6) synthesizing SA-GQDs @ ZIF-8;
    (7) synthesizing Ab2-SA-GQDs @ ZIF-8;
    (8) construction of a photoelectric sensor (PEC);
    (9) electrochemical detection of the photosensor.
  2. 2. The method for synthesizing Bi according to the construction of GQDs @ ZIF-8 as a signal quencher sensor in claim 14NbO8Cl, whichIs characterized in that: simply mix 1.3 g of BiOCl, 0.66 g of Nb 2O53.49 g of Bi2Grinding O, 2.92 g of NaCl and 3.7 g of KCl powder for 10 min, then calcining the uniformly mixed powder at 700 ℃ for 3 h, washing the powder with water at 90 ℃ for several times, and drying and collecting the powder.
  3. 3. The construction of GQDs @ ZIF-8 as a signal quencher sensor according to claim 1, synthesizing Bi4NbO8Cl/SnIn4S8A heterojunction, characterized by: 0.2 g of SnCl4·5H2O, 0.70 g of InCl3·4H2O and 0.45 g of thioacetamide in 70 mL of absolute ethanol; then 0.05 g of the powder synthesized in step (1) was dispersed in the above solution, heated to 70 ℃ by an oil bath and kept for 3 hours, and finally the resulting precipitate was centrifuged and washed 5 times with ethanol; finally, the product obtained is dried at 80 ℃ overnight to obtain Bi4NbO8Cl/SnIn4S8A heterojunction.
  4. 4. The GQDs @ ZIF-8 sensor as claimed in claim 1, constructed as ITO/Bi4NbO8Cl/SnIn4S8An 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; bi synthesized in the step (2) and having a concentration of 2.0 mg/mL4NbO8Cl/SnIn4S8Dripping on ITO glass, drying at 60 deg.C to obtain ITO/Bi 4NbO8Cl/SnIn4S8And an electrode.
  5. 5. The construction of GQDs @ ZIF-8 as a signal quencher sensor according to claim 1, which synthesizes GQDs, and is characterized in that:
    a mixture of 1.0 g glucose, 400. mu.L ethylenediamine and 200. mu.L HCl (37%, w/w) was added to 15 mL deionized water and stirred for 30 min; conveying the reaction mixture into an autoclave, and carrying out a hydrothermal process for 6 hours at the temperature of 200 ℃; the brown product from this step was centrifuged and dialyzed to remove large pieces of graphene oxide and the resulting GQDs solution was stored at 4 ℃.
  6. 6. The GQDs @ ZIF-8 as defined in claim 1 is synthesized as GQDs @ ZIF-8 by construction of a signal quencher sensor, wherein: 1.485 g of Zn (NO)3)2·6H2O and 3.28 g of 2-methylimidazole are dissolved in 50 mL of methanol respectively; 5 mL of the solution of step (4) was mixed with Zn (NO)3)2·6H2Mixing the O solution for 0.5 h under magnetic stirring, and quickly pouring into the mixture; then stirring for 2 hours at room temperature; finally, the product was centrifuged, washed with methanol and dried overnight.
  7. 7. The method for constructing the GQDs @ ZIF-8 sensor as claimed in claim 1, wherein the synthetic SA-GQDs @ ZIF-8 is characterized in that: adding 0.5 mL of 1 mg/mL Streptavidin (SA) to 2 mL of the product synthesized in step (5), stirring the mixture vigorously for 1 min and after 12 h of gentle shaking at 4 ℃, SA-labeled GQDs @ ZIF-8 are obtained; and stored at 4 ℃ protected from light for future use.
  8. 8. The method for synthesizing Ab2-SA-GQDs @ ZIF-8 by using GQDs @ ZIF-8 as a signal quencher sensor as claimed in claim 1, wherein the method comprises the following steps: and (3) 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, washing with phosphate buffer solution with the pH of 7.4 for 3 times to remove uncomplexed Ab2, and thus obtaining Ab2-SA-GQDs @ ZIF-8.
  9. 9. The construction of GQDs @ ZIF-8 as a signal quencher sensor, a photoelectric sensor (PEC), according to claim 1, wherein: rinsing ITO/Bi with ultrapure water4NbO8Cl/SnIn4S8Electrodes, 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 dropping and coating 20 mu L3% bovine serum albumin sealBlocking non-specific binding sites, thoroughly washing with phosphate buffer solution with pH 7.4 for 3 times, dripping 20 μ L of prostate antigen with different concentrations onto the surface of the electrode, incubating at room temperature for 30 min, and washing with phosphate buffer solution with pH 7.4 for 3 times; continuing to add 20. mu.L of the synthesized product of step (7), incubating at room temperature for 4H, and incubating the modified electrode in a solution containing 1 mM H2O2Incubated for 20 min in 10 mM 4 chloro-1-naphthol solution.
  10. 10. The GQDs @ ZIF-8 as claimed in claim 1, which is used for construction of a signal quencher sensor and electrochemical detection of a photoelectric sensor, and is characterized in that: and (3) taking the modified electrode processed in the step (8) 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 a phosphate buffer solution system (1 mol/L ascorbic acid) with the pH value of 7.4 as an electrolytic cell, and measuring a current I-T curve to detect the photoelectric property.
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