CN114674894B - Construction of GQDs@ZIF-8 as a signal quencher sensor - Google Patents
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Abstract
The invention discloses a construction method of a sensor based on GQDs@ZIF-8 as a signal quencher, which synthesizes Bi at first 4 NbO 8 Cl/SnIn 4 S 8 Heterojunction wherein Bi 4 NbO 8 Cl is a silicon-based perovskite, has visible light absorption property, good stability and water resistance, and [ Bi ] 2 O 2 ]And [ NbO ] 4 ]The layered structure of the layer sequence is such that Bi 4 NbO 8 The carriers of Cl can migrate rapidly; construction of Bi because of the appropriate band position 4 NbO 8 Cl/SnIn 4 S 8 The 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 steric hindrance effect 4 NbO 8 Cl/SnIn 4 S 8 The photocurrent signal of the electrode can also be used as a simulator 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
Technical Field
The invention relates to the field of quantitative detection of prostate specific antigen, in particular to construction of GQDs@ZIF-8 serving as a sensor of a signal quencher.
Background
Along with the continuous deterioration of the environment, the probability of suffering from cancer is increased, and the existing modes such as radiotherapy, chemotherapy and the like can generate serious damage to normal cells while treating the cancer, so that the early detection and treatment of the cancer have very important significance for diagnosing malignant tumors, and the establishment of a novel method for detecting malignant tumor biomarkers with simplicity, rapidness, sensitivity and good selectivity has very important value for early detection and treatment effect evaluation of the malignant tumors.
Photoelectrochemistry has attracted extensive research as a promising analytical method. Compared with the traditional electrochemistry, the PEC has the advantages of high sensitivity, good selectivity and low determination cost due to the separation of an excitation light source and an output signal. Therefore, the method is widely applied to the fields of disease diagnosis, food safety detection, environmental protection and the like due to high sensitivity. Photoelectrochemical immunosensor outputs a signal expressed by a sensitive biological material as an electric signal by immobilizing the sensitive biological material such as an active substance of enzyme, antigen, antibody, DNA, etc. as a recognition element. The specificity recognition of the biological material enables the photoelectrochemical immunosensor to have good specificity and sensitivity to 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 problems, the invention is realized by the following measures: the construction of GQDs@ZIF-8 as a signal quencher sensor is characterized by comprising the following steps:
(1) Synthesis of Bi 4 NbO 8 Cl:1.3 g BiOCl, 0.66 g Nb 2 O 5 Bi of 3.49. 3.49 g 2 O, 2.92 and g NaCl and 3.7 and g 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 Bi 4 NbO 8 Cl/SnIn 4 S 8 Heterojunction: 0.2 g SnCl 4 ·5H 2 O, inCl of 0.70 g 3 ·4H 2 O and 0.45. 0.45 g thioacetamide are dissolved in 70 mL absolute ethanol; then dispersing 0.05 g of the powder synthesized in the step (1) in the solution, heating to 70 ℃ through an oil bath and keeping the temperature at 3 h, and finally centrifuging the obtained precipitate and washing the precipitate with ethanol for 5 times; finally, the obtained product was dried overnight at 80 ℃ to obtain Bi 4 NbO 8 Cl/SnIn 4 S 8 And a heterojunction.
(3) Construction of ITO/Bi 4 NbO 8 Cl/SnIn 4 S 8 An electrode: the conductive glass is indium tin oxide glass (ITO), the conductive glass is cut into 4.0x0.5 cm strips, sequentially washed by acetone solution, secondary distilled water and absolute ethyl alcohol for 5 min in an ultrasonic manner, and then dried under nitrogen for standby; bi synthesized in the step (2) is synthesized to have the concentration of 2.0 mg/mL 4 NbO 8 Cl/SnIn 4 S 8 Dripping onto ITO glass, and drying at 60deg.C to obtain ITO/Bi 4 NbO 8 Cl/SnIn 4 S 8 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; the reaction mixture was fed to an autoclave and a hydrothermal process of 6 h was performed at 200 ℃; the brown product from this step was centrifuged and dialyzed to remove large pieces of graphene oxide, and the final GQDs solution was stored at 4 ℃.
(5) Synthesis of GQDs@ZIF-8:1.485 g Zn (NO) 3 ) 2 ·6H 2 O and 3.28 g of 2-methylimidazole were dissolved in 50 mL methanol, respectively; combining the 5 mL step (4) solution with Zn (NO) 3 ) 2 ·6H 2 Mixing O solution under magnetic stirring for 0.5. 0.5 h, and rapidly pouring into the mixture; then stirring 2 h at room temperature; finally, the resulting product was centrifuged, washed with methanol and dried overnight.
(6) Synthesizing SA-GQDs@ZIF-8: 1 mg/mL of Streptavidin (SA) of 0.5. 0.5 mL was added to 1 mg/mL of the product synthesized in step (5) of 2 mL, and after vigorously stirring the mixture for 1 min and gently shaking 12 h at 4 ℃, SA-labeled GQDs@ZIF-8 was obtained; and stored in the dark at 4 ℃ for future use.
(7) Ab2-SA-GQDs@ZIF-8 was synthesized: adding the secondary antibody, namely Ab2, with the concentration of 1 mL of 10 mug/mL into the synthesized product in the step (6), incubating 2 h at the temperature of 4 ℃, and washing 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): rinsing ITO/Bi with ultra pure water 4 NbO 8 Cl/SnIn 4 S 8 The electrode was then incubated 16 h with 6 μl of primary antibody, ab1, at a concentration of 10 μg/mL at 4 ℃ and thoroughly rinsed 3 times with phosphate buffer pH 7.4; continuously dripping 20 mu L of 3% bovine serum albumin to block the non-specific binding site, thoroughly flushing 3 times by using phosphate buffer solution with the pH of 7.4, dripping 20 mu L of prostate antigens with different concentrations onto the surface of an electrode, incubating for 30 min at room temperature, and washing 3 times by using the phosphate buffer solution with the pH of 7.4; continue to drop 20. Mu.L of the product synthesized in step (7), incubate 4H at room temperature, and add the modified electrode in the presence of 1 mM H 2 O 2 Is incubated for 20 min with 10 mM 4-chloro-1-naphthol solution.
(9) Electrochemical detection by a photoelectric sensor: and (3) taking the modified electrode treated in the step (8) as a working electrode, taking a platinum wire electrode as a counter electrode, taking an Ag/AgCl electrode as a reference electrode, taking a bias voltage value of 0V, taking a xenon lamp as a light source for stimulation, taking a phosphate buffer solution system (1 mol/L ascorbic acid) with pH of 7.4 as an electrolytic cell, and measuring a current I-T curve to detect photoelectric performance.
The invention has the beneficial effects that:
（1）Bi 4 NbO 8 cl is taken as a silicon-based perovskite, has visible light absorption performance, and has good stability and water resistance.
（2）[Bi 2 O 2 ]And [ NbO ] 4 ]The layered structure of the layer sequence is such that Bi 4 NbO 8 The carriers of Cl can migrate rapidly.
(3) Construction of Bi because of the appropriate band position 4 NbO 8 Cl/SnIn 4 S 8 The heterojunction can effectively improve the charge separation efficiency.
(4) GQDs@ZIF-8 polyhedron serving as signal quencher can inhibit ITO/Bi through steric hindrance effect 4 NbO 8 Cl/SnIn 4 S 8 The photocurrent signal of the electrode can also be used as a simulator 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
For further understanding of the present invention, embodiments are provided by combining the technical solutions of the present invention with examples:
(1) Synthesis of Bi 4 NbO 8 Cl:1.3 g BiOCl, 0.66 g Nb 2 O 5 Bi of 3.49. 3.49 g 2 O, 2.92 and g NaCl and 3.7 and g 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 Bi 4 NbO 8 Cl/SnIn 4 S 8 Heterojunction: 0.2 g SnCl 4 ·5H 2 O, inCl of 0.70 g 3 ·4H 2 O and 0.45. 0.45 g thioacetamide are dissolved in 70 mL absolute ethanol; then dispersing 0.05 g of the powder synthesized in the step (1) in the solution, heating to 70 ℃ through an oil bath and keeping the temperature at 3 h, and finally centrifuging the obtained precipitate and washing the precipitate with ethanol for 5 times; finally, the obtained product was dried overnight at 80 ℃ to obtain Bi 4 NbO 8 Cl/SnIn 4 S 8 And a heterojunction.
(3) Construction of ITO/Bi 4 NbO 8 Cl/SnIn 4 S 8 An electrode: the conductive glass is indium tin oxide glass (ITO), the conductive glass is cut into 4.0x0.5 cm strips, sequentially washed by acetone solution, secondary distilled water and absolute ethyl alcohol for 5 min in an ultrasonic manner, and then dried under nitrogen for standby; bi synthesized in the step (2) is synthesized to have the concentration of 2.0 mg/mL 4 NbO 8 Cl/SnIn 4 S 8 Dripping onto ITO glass, and drying at 60deg.C to obtain ITO/Bi 4 NbO 8 Cl/SnIn 4 S 8 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; the reaction mixture was fed to an autoclave and a hydrothermal process of 6 h was performed at 200 ℃; the brown product from this step was centrifuged and dialyzed to remove large pieces of graphene oxide, and the final GQDs solution was stored at 4 ℃.
(5) Synthesis of GQDs@ZIF-8:1.485 g Zn (NO) 3 ) 2 ·6H 2 O and 3.28 g of 2-methylimidazole were dissolved in 50 mL methanol, respectively; combining the 5 mL step (4) solution with Zn (NO) 3 ) 2 ·6H 2 Mixing O solution under magnetic stirring for 0.5. 0.5 h, and rapidly pouring into the mixture; then stirring 2 h at room temperature; finally, the resulting product was centrifuged, washed with methanol and dried overnight.
(6) Synthesizing SA-GQDs@ZIF-8: 1 mg/mL of Streptavidin (SA) of 0.5. 0.5 mL was added to 1 mg/mL of the product synthesized in step (5) of 2 mL, and after vigorously stirring the mixture for 1 min and gently shaking 12 h at 4 ℃, SA-labeled GQDs@ZIF-8 was obtained; and stored in the dark at 4 ℃ for future use.
(7) Ab2-SA-GQDs@ZIF-8 was synthesized: adding the secondary antibody, namely Ab2, with the concentration of 1 mL of 10 mug/mL into the synthesized product in the step (6), incubating 2 h at the temperature of 4 ℃, and washing 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 Photosensor (PEC): rinsing ITO/Bi with ultra pure water 4 NbO 8 Cl/SnIn 4 S 8 The electrode was then incubated 16 h with 6 μl of primary antibody, ab1, at a concentration of 10 μg/mL at 4 ℃ and thoroughly rinsed 3 times with phosphate buffer pH 7.4; continuously dripping 20 mu L of 3% bovine serum albumin to block the non-specific binding site, thoroughly flushing 3 times by using phosphate buffer solution with the pH of 7.4, dripping 20 mu L of prostate antigens with different concentrations onto the surface of an electrode, incubating for 30 min at room temperature, and washing 3 times by using the phosphate buffer solution with the pH of 7.4; continue to drop 20. Mu.L of the product synthesized in step (7), incubate 4H at room temperature, and add the modified electrode in the presence of 1 mM H 2 O 2 Is incubated for 20 min with 10 mM 4-chloro-1-naphthol solution.
(9) Electrochemical detection by a photoelectric sensor: the modified electrode treated in the step (8) is used as a working electrode, a counter electrode is a platinum wire electrode, a reference electrode is an Ag/AgCl electrode, the bias value is 0V, a xenon lamp is used as a light source for stimulation, an electrolytic cell is a phosphate buffer system (1 mol/L ascorbic acid) with pH of 7.4, and the photoelectric performance is detected by measuring a current I-T curve, so that a linear equation I= -1.23log (c) -10.24, the correlation coefficient is 0.994, the detection limit is 0.07 pg/mL, and the high-sensitivity detection of the prostate specific antigen is realized.
Claims (1)
1. The construction method of GQDs@ZIF-8 serving as a signal quenching sensor is characterized by comprising the following steps of:
(1) Synthesis of Bi 4 NbO 8 Cl:1.3 g BiOCl, nb 0.66. 0.66 g 2 O 5 Bi of 3.49. 3.49 g 2 Grinding O, 2.92 and g NaCl and 3.7 and g KCl powder for 10 min, calcining the uniformly mixed powder at 700 ℃ for 3 h, washing with water at 90 ℃ for several times, and drying and collecting;
(2) Synthesis of Bi 4 NbO 8 Cl/SnIn 4 S 8 Heterojunction: 0.2 g SnCl 4 ·5H 2 O, inCl of 0.70 g 3 ·4H 2 O and 0.45. 0.45 g thioacetamide are dissolved in 70 mL absolute ethanol to form a mixed solution; then dispersing 0.05 g of the powder synthesized in the step (1) in the solution, heating to 70 ℃ through an oil bath and keeping the temperature at 3 h, and finally centrifuging the obtained precipitate and washing the precipitate with ethanol for 5 times; finally, the obtained product was dried overnight at 80 ℃ to obtain Bi 4 NbO 8 Cl/SnIn 4 S 8 A heterojunction;
(3) Construction of ITO/Bi 4 NbO 8 Cl/SnIn 4 S 8 An electrode: the conductive glass is indium tin oxide glass (ITO), the conductive glass is cut into 4.0x0.5 cm strips, sequentially washed by acetone solution, secondary distilled water and absolute ethyl alcohol for 5 min in an ultrasonic manner, and then dried under nitrogen for standby; bi synthesized in the step (2) is synthesized to have the concentration of 2.0 mg/mL 4 NbO 8 Cl/SnIn 4 S 8 Dripping onto ITO glass, and drying at 60deg.C to obtain ITO/Bi 4 NbO 8 Cl/SnIn 4 S 8 An electrode;
(4) Synthesis of GQDs: a mixture of 1.0 g glucose, 400 μl ethylenediamine and 200 μl concentrated HCl was added to 15 mL deionized water and stirred for 30 min; the reaction mixture was fed to an autoclave and subjected to a hydrothermal process of 6 h at 200 ℃; centrifuging and dialyzing the brown product obtained from the step to remove large graphene oxide, and storing the GQDs solution obtained finally at 4 ℃;
(5) Synthesis of GQDs@ZIF-8:1.485 g Zn (NO) 3 ) 2 ·6H 2 O and 3.28 g of 2-methylimidazole were dissolved in 50 mL methanol, respectively; combining the 5 mL step (4) solution with Zn (NO) 3 ) 2 ·6H 2 Mixing O solution under magnetic stirring for 0.5. 0.5 h, and rapidly pouring into the mixture; then stirring 2 h at room temperature; finally, centrifuging the obtained product, washing with methanol, and drying overnight;
(6) Synthesizing SA-GQDs@ZIF-8: 1 mg/mL of Streptavidin (SA) of 0.5. 0.5 mL was added to 1 mg/mL of the product synthesized in step (5) of 2 mL, and after vigorously stirring the mixture for 1 min and gently shaking 12 h at 4 ℃, SA-labeled GQDs@ZIF-8 was obtained; and stored in a dark place at 4 ℃ for future use;
(7) Ab2-SA-GQDs@ZIF-8 was synthesized: adding the secondary antibody, namely Ab2, with the concentration of 1 mL of 10 mug/mL into the synthesized product of the step (6), incubating 2 h at the temperature of 4 ℃, and washing 3 times with 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 Photosensor (PEC): rinsing ITO/Bi with ultra pure water 4 NbO 8 Cl/SnIn 4 S 8 The electrode was then incubated 16 h with 6 μl of primary antibody, ab1, at a concentration of 10 μg/mL at 4 ℃ and thoroughly rinsed 3 times with phosphate buffer pH 7.4; continuously dripping 20 mu L of 3% bovine serum albumin to block the non-specific binding site, thoroughly flushing 3 times by using phosphate buffer solution with the pH of 7.4, dripping 20 mu L of prostate antigens with different concentrations onto the surface of an electrode, incubating for 30 min at room temperature, and washing 3 times by using the phosphate buffer solution with the pH of 7.4; continue to drop 20. Mu.L of the product synthesized in step (7), incubate 4H at room temperature, and add the modified electrode in the presence of 1 mM H 2 O 2 Is incubated for 20 min with 10 mM 4-chloro-1-naphthol solution.
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| WO2018094779A1 (en) * | 2016-11-25 | 2018-05-31 | 深圳大学 | Photoelectrochemical biosensor and preparation method therefor |
| CN108845119A (en) * | 2018-04-13 | 2018-11-20 | 济南大学 | A kind of construction method of the photic electrochemical immunosensor based on carrier double regulation control strategy |
| CN113984854A (en) * | 2021-10-18 | 2022-01-28 | 济南大学 | Preparation method of aflatoxin B1 photoelectrochemical sensor constructed based on double Z-type heterojunction |
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| WO2018094779A1 (en) * | 2016-11-25 | 2018-05-31 | 深圳大学 | Photoelectrochemical biosensor and preparation method therefor |
| CN108845119A (en) * | 2018-04-13 | 2018-11-20 | 济南大学 | A kind of construction method of the photic electrochemical immunosensor based on carrier double regulation control strategy |
| CN113984854A (en) * | 2021-10-18 | 2022-01-28 | 济南大学 | Preparation method of aflatoxin B1 photoelectrochemical sensor constructed based on double Z-type heterojunction |
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| Visible-light-driven double-shell SnIn4S8/TiO2 heterostructure with enhanced photocatalytic activity for MO removal and Cr(VI) cleanup;Jun Wang 等;《Applied Surface Science》;第152867（1-14）页 * |
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