CN112858417A - Photoelectrochemical sensor detection m based on bismuth sulfide-silver bromide heterojunction6Method A - Google Patents
Photoelectrochemical sensor detection m based on bismuth sulfide-silver bromide heterojunction6Method A Download PDFInfo
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Abstract
The invention discloses a method for preparing Bi based on flower shape2S3Photoelectrochemical analysis of (A) to detect N in RNA6The method of-methyl adenine ribonucleotide, firstly constructing the method for detecting N6-a photoelectrochemical biosensor of methyladenine ribonucleotides comprising: an electrode, flower-like Bi sequentially modified on the surface of the electrode2S3Chitosan functionalized AgBr, 4-carboxyphenylboronic acid and N6-methyladenine antibody, N6-methyladenine ribonucleotide, CeO2. The invention utilizes Bi2S3Photoelectric property of heterojunction material formed by AgBr and specificity recognition of antibody to antigenCan, and CeO2The nano material is used for signal amplification to realize N in RNA6Detection of methyladenine ribonucleotides. The method has simple detection method, realizes instrument miniaturization, is easy to operate, and can realize N in RNA only by simply modifying the surface of the ITO electrode6Detection of methyladenine ribonucleotides.
Description
Technical Field
The invention relates to the technical field of photoelectrochemical analysis, in particular to a flower-shaped Bi-based material2S3Photoelectrochemical analytical detection of N by AgBr heterojunction6-methyladenine ribonucleotides.
Background
RNA methylation is a very important part of the field of epigenetic modification. At the RNA level, N6-methyladenine ribonucleotides are the most predominant methylation-modified form of RNA. In mammals, N6-methyladenine ribonucleotide modification in a proportion of 0.1 to 0.4%, with an average of 3 to 5N per mRNA6-methyladenine ribonucleotide sites. N is a radical of6-methyladenine ribonucleotides play an important role in a variety of cancer types, including leukemia, brain tumors, liver cancer, breast cancer and lung cancer. The research shows that N6Dynamic regulation of methyladenine ribonucleotides has a significant impact on the control of gene expression. Various studies have demonstrated that N6Importance of-methyladenine ribonucleotides, but analysis and evaluation of N due to lack of sensitive and highly selective analytical methods6The effect of the-methyladenine ribonucleotide content on the study was greatly influenced, thus achieving an effect on N6The detection of-methyladenine ribonucleotide and the content thereof has important significance.
Currently for N6The related detection method of-methyladenine ribonucleotide mainly comprises thin layer chromatography, gas chromatography, column liquid chromatography, liquid chromatography-mass spectrometry combined method and capillary electrophoresis method. Although the above method is for detecting N6-methyladenine ribonucleotide (m)6A) Has great effect, but also has some disadvantages, such as the need32P labeling or other radioactive element labeling, the need of precise and complex instruments, complex sample pretreatment, the need of professional operators, low detection sensitivity, weak specificity and the like. Thus, a simple, fast, highly sensitive and highly selective method for N is established6The detection of methyladenine ribonucleotides is of crucial importance.
The photoelectrochemical sensor is a novel analytical detection technique based on the light-to-electricity conversion characteristic of the photoactive material, and has the advantages of electrochemical analysis and photochemical analysis. Which excites the electro-optically active material with light to produce photo-generated electrons and holes. The photo-generated electrons are captured by the electrodes to generate an electric current. The excitation light source and the detection signal are in two completely different forms, so that the interference of the background signal can be effectively reduced, the sensitivity of analysis and detection is greatly improved, and the method has the advantages of high detection speed, easiness in miniaturization, simplicity in operation, simplicity in instrument, low background signal and the like. At present, no flower-shaped Bi-based material exists2S3Photoelectrochemical analytical detection of N by AgBr heterojunction6-a report of methyladenine ribonucleotides.
Disclosure of Invention
In view of the above prior art, the present invention provides a method for detecting N6The photoelectrochemical sensor of the-methyladenine ribonucleotide and the detection method thereof realize the N pair6High specificity and high sensitivity detection of methyladenine ribonucleotides.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect of the invention, a method for detecting N is provided6-a photoelectrochemical biosensor of methyladenine ribonucleotides comprising an electrode; the surface of the electrode is sequentially modified with flower-shaped Bi2S3AgBr, 4-carboxyphenylboronic acid, N6-methyladenine antibody, N6-methyladenine ribonucleotides and CeO2。
Preferably, the electrode is an ITO electrode.
In a second aspect of the present invention, there is provided a method for preparing the above photoelectrochemical biosensor, comprising the steps of:
(1) flower-shaped Bi2S3Fixed on a pretreated electrode to obtain Bi2S3An electrode;
(2) AgBr is fixed on Bi obtained in the step (1)2S3Per electrode surface to obtain AgBr/Bi2S3An electrode;
(3) modifying 4-carboxyphenylboronic acid (CPBA) to AgBr/Bi obtained in step (2)2S3Per electrode surface to obtain CPBA/AgBr/Bi2S3An electrode;
(4) will N6Modification of the CPBA/AgBr/Bi obtained in step (3) with a methyladenine antibody2S3To obtain Ab/CPBA/AgBr/Bi on the electrode surface2S3An electrode;
(5) using specific binding between antigen and antibody to convert N6Modification of methyl adenine ribonucleotide to Ab/CPBA/AgBr/Bi obtained in step (4)2S3Per electrode surface, to obtain m6A/Ab/CPBA/AgBr/Bi2S3An electrode;
(6) CeO by utilizing specific combination between phosphate radical and cerium2Modifying m obtained in step (5)6A/Ab/CPBA/AgBr/Bi2S3On the surface of an electrode to obtain CeO2/m6A/Ab/CPBA/AgBr/Bi2S3Electrode, i.e. detecting N6-a photoelectrochemical biosensor of methyladenine ribonucleotides.
Preferably, in the step (1), the method for pretreating the electrode comprises the following steps: the electrodes were treated with acetone and 1-4M NaOH in aqueous alcohol (V)Anhydrous ethanol:VSecondary water1:1-1:6) and secondary water are respectively ultrasonically cleaned for 15-40min and dried.
Preferably, in step (1), Bi is flower-shaped2S3The fixing method comprises the following steps:
flower-shaped Bi2S3The nano material is evenly dispersed in deionized water to obtain Bi2S3A dispersion of Bi2S3Dripping the dispersed liquid on the surface of a pretreatment electrode, and drying under the irradiation of an infrared lamp;
more preferably, the flower-like Bi2S3The nano material is prepared by the following method:
separately adding Bi (NO)3)3·5H2Dissolving O, thiourea, urea and 4-carboxyphenylboronic acid in ethylene glycol, carrying out hydrothermal reaction, collecting precipitate, centrifugally washing with water and ethanol, drying, and collecting solid to obtain flower-shaped Bi2S3And (3) nano materials.
Preferably, in the step (2), the AgBr immobilization method comprises:
uniformly dispersing AgBr nano materials in deionized water to obtain AgBr dispersion liquid, dropwise adding the AgBr dispersion liquid on the electrode treated in the step (1), and drying under the irradiation of an infrared lamp;
more preferably, the AgBr nano material is prepared by the following method:
mixing AgNO3And mixing the aqueous solution and the chitosan aqueous solution, adding the KBr aqueous solution in the stirring process, stirring for 1-5h to form a dispersion containing AgBr nano particles, centrifugally collecting precipitates, centrifugally washing with water and ethanol, drying and collecting solids to obtain the AgBr nano material.
Preferably, in the step (3), the method for modifying the 4-carboxyphenylboronic acid is as follows:
dropwise adding the activated 4-carboxyphenylboronic acid solution onto the electrode treated in the step (2), reacting for 0.5-3h under the humid condition of 37 ℃, and then cleaning for 1-5 times;
more preferably, the activated 4-carboxyphenylboronic acid solution is prepared by:
respectively adding 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) solutions into a 4-carboxyphenylboronic acid solution, and activating at 37 ℃ for 10-90min to obtain the activated 4-carboxyphenylboronic acid solution; the 4-carboxyphenylboronic acid solution is prepared by PBS buffer solution with pH of 5.5-8.5 and concentration of 5-500mM, and the concentration of the 4-carboxyphenylboronic acid solution is 0.5-5 mM; the EDC solution and the NHS solution are respectively prepared by PBS buffer solution with pH of 5.5-8.5 and concentration of 5-500mM, and the concentration of the EDC solution and the concentration of the NHS solution are both 0.5 mg/mL.
Preferably, in step (4), N is6The method for modifying the-methyladenine antibody comprises the following steps:
will N6Dropwise adding a methyl adenine antibody solution onto the electrode treated in the step (3), reacting for 0.5-3h under the humid condition of 37 ℃, and then washing for 1-5 times.
Preferably, in step (5), N is6The method for modifying the-methyladenine ribonucleotide comprises the following steps:
will N6Dropwise adding a methyl adenine ribonucleotide solution onto the surface of the electrode treated in the step (4), reacting for 0.5-3h under the humidity condition of 37 ℃, and then cleaning for 1-5 times.
Preferably, in the step (6), CeO2The modification method comprises the following steps:
adding CeO2Dropwise adding the nano material dispersion liquid onto the electrode treated in the step (5), reacting for 0.5-5h under the humid condition of 37 ℃, and then cleaning for 1-5 times;
more preferably, the CeO2The nano material is prepared by the following method:
respectively adding Na3PO4·12H2O and Ce (NO)3)3·6H2Dissolving O in deionized water, carrying out hydrothermal reaction, collecting precipitate, centrifugally washing with water and ethanol, drying, and collecting solid to obtain CeO2And (3) nano materials.
In a third aspect of the present invention, there is provided the above-mentioned photoelectrochemical biosensor for detecting N6-methyladenine ribonucleotides.
In a fourth aspect of the present invention, there is provided the above-mentioned photoelectrochemical biosensor for detecting N6-a method of methyladenine ribonucleotides comprising the steps of:
a photoelectrochemical biosensor is used as a working electrode, a saturated calomel electrode is used as a reference electrode, a Pt wire is used as an auxiliary electrode to form a three-electrode system for photoelectrochemical signal detection, electrode detection solution is PBS buffer solution containing Ascorbic Acid (AA) (the pH of the PBS buffer solution is 5.5-8.5, and the concentration of the PBS buffer solution is 5-500mM), and current and N are established6Concentration of-methyladenine nucleoside sugar nucleotideRelation between, to N6-detecting the content of methyladenine ribonucleotides.
Preferably, for N6The method for detecting the content of the-methyladenine ribonucleotide is a current-time method, and the applied potential is as follows: -0.5-0.5V.
The invention has the beneficial effects that:
(1) the invention utilizes CeO2For Bi2S3The function of the-AgBr heterojunction realizes the amplification of photoelectric signals and improves N6Sensitivity of detection of methyladenine ribonucleotides.
(2) The present invention utilizes the specific reaction between phenylboronic acid and antibody, and N6Specific recognition of the-methyladenine antibody antigen by increasing N6Specificity of detection of methyladenine ribonucleotides.
(3) The detection method is simple, has low cost, realizes the miniaturization of instruments, and can realize the N-ray detection only by simply processing the surface of the ITO electrode6Detection of methyladenine ribonucleotides.
Drawings
FIG. 1: photoelectrochemical biosensor construction of the invention and N6Schematic diagram of the detection of methyladenine ribonucleotides.
FIG. 2: photocurrent intensity and N6-a linear fit curve of the logarithmic values of the concentration of methyladenine ribonucleotides.
FIG. 3: histogram of changes in photoelectrochemical response under different nucleotide conditions.
Detailed Description
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
Description of the drawings: the "room temperature" in the present invention is in the range of 20 to 30 ℃.
"humid conditions" in the present invention are humidity greater than 90%; the preferred humidity is 95-99%.
The cleaning solution used in the invention comprises the following components: 3-15mM Tris-HCl and 20-60mM KCl, pH 7.4.
N used in the present invention6The components of the-methyladenine antibody buffer solution are as follows: 5-30mM PBS, pH 6.0-8.5.
The RNA lysis solution used in the present invention comprises the following components: 5-100mM Tris, 1-50mM Ethylene Diamine Tetraacetic Acid (EDTA), pH 5.0-8.5, the solvent is sterilized water (the sterilized water in this invention is DEPC treated water).
As described in the background section, the prior art is capable of detecting N6-methyladenine ribonucleotides, but also suffer from disadvantages such as the need for32P labeling or other radioactive element labeling, the need of precise and complex instruments, complex sample pretreatment, the need of professional operators, low detection sensitivity, weak specificity and the like.
Based on the above, the invention constructs a method for detecting N6The principle diagram of the construction and detection of the photoelectrochemical biosensor of the present invention is shown in FIG. 1. The photoelectrochemical biosensor takes an ITO electrode as a substrate electrode and takes flower-shaped Bi2S3And AgBr are sequentially modified on the surface of the electrode. The CPBA is modified on the surface of the electrode by utilizing the action between the amino on the AgBr surface after the chitosan functionalization and the carboxyl of 4-carboxyl phenylboronic acid (CPBA). By utilizing the specific reaction of the boric acid group and the antibody, N is reacted6-methyladenine antibody is modified to the electrode surface. By specific recognition of antigen-antibody, N6-methyladenine ribonucleotides are modified onto the electrode surface. Utilizing the specific combination between phosphate radical and cerium ion to combine CeO2And modifying the surface of the electrode. Wherein, the flower-shaped Bi2S3Is an excellent photoelectric material, AgBr and Bi2S3Heterojunction is formed, photocurrent is increased, the surface of the electrode is provided with amino, 4-carboxyl phenylboronic acid is modified on the surface of the electrode, and N is further modified6-methyladenine antibody, thereby introducing N6-methyladenine ribonucleotides, blocking the migration of photo-generated electrons, reducing the photocurrent, and mostPost-introduction of CeO2As a signal amplification molecule, the transfer of interface electrons is accelerated, and signal amplification is realized. And CeO2Modified by the amount of N6-concentration of methyladenine ribonucleotides. Thus, using N6The linear relation between the concentration of the-methyladenine ribonucleotide and the intensity of photocurrent can be realized for N6-methyladenine ribonucleotide (m)6A) Detection of (2), N6-methyladenine ribonucleotide (m)6A) The structural formula of (A) is as follows:
in one embodiment of the present invention, the construction process of the photoelectrochemical biosensor is as follows:
(1) flower-like Bi2S3The preparation of (1): respectively adding 1.0-3.0g of Bi (NO)3)3·5H2Dissolving O, 0.2-1.0g of thiourea, 1.0-3.0g of urea and 0.01-0.1g of cetyltrimethylammonium bromide (CTAB) in 25, 30, 20 and 10mL of ethylene glycol respectively, mixing the solutions under stirring, stirring for 10-60min, transferring the mixture into a reaction kettle, standing at the temperature of 150-2S3And (3) nano materials.
(2) Preparation of chitosan functionalized AgBr: 20mL of 0.05-0.2M AgNO3Mixing the aqueous solution with 20 mL0.1-1.0% (M/v) chitosan aqueous solution, stirring for 10-60min, adding 20mL of 0.1-0.5M KBr aqueous solution during stirring, continuously stirring for 1-5h to form a dispersion containing AgBr nanoparticles, centrifuging at 10000-12000rpm for 10min to collect precipitates, centrifuging and washing for 2-6 times by using water and absolute ethyl alcohol, drying at 40-60 ℃ and collecting solids to obtain the chitosan functionalized AgBr nano material. All the above procedures were carried out in the dark.
(3)CeO2The preparation of (1): mixing 0.01-0.05gNa3PO4·12H2Dissolving O in 105mL deionized water, stirring for 10min, and stirringSlowly adding 15mL of 0.1-0.5M Ce (NO)3)3·6H2O water solution, transferring the mixture into a reaction kettle, standing at the temperature of 150-2And (3) nano materials.
(4)Bi2S3Preparation of the dispersion: weighing 2-10mg of flower-shaped Bi prepared in the step (1)2S3Adding the nano material into 2-10mL of deionized water, and performing ultrasonic dispersion for 0.5-3 hours.
(5) Preparation of AgBr dispersion: and (3) weighing 2-10mg of the chitosan functionalized AgBr nano material prepared in the step (2), adding the chitosan functionalized AgBr nano material into 2-10mL of deionized water, and performing ultrasonic dispersion for 0.5-3 hours.
(6)CeO2Preparation of the dispersion: weighing 2-10mg of CeO prepared in the step (3)2Adding into 2-10mL deionized water, and ultrasonic dispersing for 0.5-3 h.
(7) Preparation of electrode washing buffer: mixing 3-15mM Tris-HCl and 20-60mM KCl with sterilized water as solvent, and adjusting pH to 7.4.
(8) Preparation of 4-carboxyphenylboronic acid solution: PBS buffer solution with pH of 5.5-8.5 and concentration of 5-500mM is prepared by using sterilized water, and 4-carboxyphenylboronic acid solution with 0.5-5mM is prepared by using the PBS buffer solution as a solvent.
(9) Preparation of EDC and NHS solutions: preparing PBS buffer solution with pH of 5.5-8.5 and concentration of 5-500mM with sterilized water, and preparing 0.5mg/mL PBS buffer solution as solvent-1EDC, NHS solution (c).
(10) Activation of 4-carboxyphenylboronic acid solution: respectively adding 10 mu L of EDC and NHS solution into 20 mu L of 4-carboxyphenylboronic acid solution, and activating at 37 ℃ for 10-90min to obtain the activated 4-carboxyphenylboronic acid solution.
(11) Pretreating an ITO electrode: cutting ITO conductive glass into 5 × 1cm2Separately using acetone and 1-4M NaOH in an aqueous alcohol solution (V)Anhydrous ethanol:VSecondary water1:1-1:6) and secondary water, carrying out ultrasonic treatment on the ITO electrode for 15-60min, then washing with the secondary water, and naturally drying for later use.
(12) Flower-like Bi2S3Fixing: 20-80 mu L of Bi2S3And dropwise adding the dispersion liquid to the surface of the pretreated ITO electrode, and irradiating and drying by using an infrared lamp. Then, the electrode is washed 1 to 5 times with a washing solution. And (5) drying by nitrogen. The prepared electrode is marked as Bi2S3/ITO。
(13) Fixing AgBr: dropping 20-80 μ LAgBr dispersion to Bi2S3Drying the ITO electrode surface by infrared lamp irradiation. Then, the electrode is washed 1 to 5 times with a washing solution. And (5) drying by nitrogen. The prepared electrode is marked as AgBr/Bi2S3/ITO。
(14) Immobilization of 4-carboxyphenylboronic acid: 10-50 mu L of activated 4-carboxyphenylboronic acid solution is dripped into AgBr/Bi2S3The surface of the ITO electrode reacts for 0.5 to 3 hours under the humid condition of 37 ℃, and then is cleaned for 1 to 5 times. The prepared electrode is marked as CPBA/AgBr/Bi2S3/ITO。
(15)N6-immobilization of methyladenine antibodies: mixing 10-50 μ L of 5-50 μ g/mL-1m6Dropwise adding the antibody A to CPBA/AgBr/Bi2S3The surface of the ITO electrode reacts for 0.5 to 3 hours under the humid condition of 37 ℃, and then is cleaned for 1 to 5 times. The prepared electrode is marked as Ab/CPBA/AgBr/Bi2S3/ITO。
(16)N6-immobilization of methyladenine ribonucleotides: mixing 10-50 μ LN6Dropping of-methyladenine ribonucleotides to Ab/CPBA/AgBr/Bi2S3The surface of the ITO electrode reacts for 0.5 to 3 hours under the humid condition of 37 ℃, and then is cleaned for 1 to 5 times. The prepared electrode is marked as m6A/Ab/CPBA/AgBr/Bi2S3/ITO。
(17)CeO2Fixing: mixing 10-60 μ L of CeO2Dropping the dispersed liquid to m6A/Ab/CPBA/AgBr/Bi2S3The surface of the ITO electrode reacts for 0.5 to 5 hours under the humid condition of 37 ℃, and then is cleaned for 1 to 5 times. The prepared electrode was labeled as CeO2/m6A/Ab/CPBA/AgBr/Bi2S3/ITO。
In the construction process of the photoelectrochemical sensor, the steps supplement each other, the sequence is strictly limited, each step serves for the next fixed modification, and the lack of the previous step can cause the failure of the subsequent modification. The material fixed and modified on the surface of the ITO electrode can be a commercially available product or can be prepared by itself, as long as the performance meets the use requirement, and the invention is not particularly limited.
In another embodiment of the present invention, the detection of N using the above-described photoelectrochemical biosensor is provided6The process of-methyladenine ribonucleotides is:
(1) preparing an electrode detection solution: PBS buffer solution with pH of 5.5-8.5 and concentration of 5-500mM is prepared with sterilized water, then Ascorbic Acid (AA) of 5-500mM is added into the solution, and the obtained solution is used as electrode detection solution.
(2) At different concentrations of N6CeO prepared from methyladenine ribonucleotides2/m6A/Ab/CPBA/AgBr/Bi2S3The ITO electrode is used as a working electrode, the saturated calomel electrode is used as a reference electrode, the Pt filament is used as an auxiliary electrode to form a three-electrode system for carrying out photoelectrochemical signal detection, a light source is visible light, the applied potential is-0.5-0.5V, and photocurrent is recorded in electrode detection liquid.
(3) Establishing photocurrent intensity and N6-the relationship between the concentration of methyladenine ribonucleotides, the use of which for N in a sample to be tested6The content of-methyladenine ribonucleotides is detected.
With N6Increase in concentration of methyladenine ribonucleotide, CeO on the surface of the electrode2Is increased, thereby causing an increase in the photoelectric signal, according to N6The linear relation between the concentration of the-methyladenine ribonucleotide and the intensity of photocurrent can be realized for N6Detection of methyladenine ribonucleotides.
Pair of photoelectrochemical biosensors using the present invention6The detection range of-methyladenine ribonucleotide is 0.001-300nM, and the detection limit is 0.33 pM.
In order to make the technical solutions of the present application more clearly understood by those skilled in the art, the technical solutions of the present application will be described in detail below with reference to specific embodiments.
The test materials used in the examples of the present invention are all conventional in the art and commercially available.
Example 1: flower-like Bi2S3Preparation of
Respectively adding 2.0g of Bi (NO)3)3·5H2Dissolving O, 0.6g of thiourea, 2.0g of urea and 0.05g of hexadecyl trimethyl ammonium bromide (CTAB) in 25, 30, 20 and 10mL of ethylene glycol respectively, mixing the solutions under stirring, stirring for 40min, transferring the mixture into a reaction kettle, standing at 175 ℃, centrifuging at 9000rpm for 10min after standing for 30h, collecting precipitates, centrifuging and washing for 4 times by using water and absolute ethyl alcohol, drying at 50 ℃ and collecting solids, wherein the obtained solids are flower-shaped Bi2S3And (3) nano materials.
Example 2: preparation of chitosan functionalized AgBr
20mL of 0.13M AgNO3Mixing the aqueous solution with 20mL of 0.5% (M/v) chitosan aqueous solution, stirring for 40min, adding 20mL of 0.3M KBr aqueous solution in the stirring process, continuously stirring for 3h to form a dispersion containing AgBr nanoparticles, centrifuging at 11000rpm for 10min, collecting precipitates, centrifuging and washing for 4 times by using water and absolute ethyl alcohol, drying at 50 ℃ and collecting solids to obtain the chitosan functionalized AgBr nano material. All the above procedures were carried out in the dark.
Example 3: CeO (CeO)2Preparation of
Adding 0.03g of Na3PO4·12H2Dissolving O in 105mL deionized water, stirring for 10min, and slowly adding 15mL 0.3M Ce (NO)3)3·6H2O water solution, transferring the mixture into a reaction kettle, standing at 175 ℃ for 30h, centrifuging at 9000rpm for 10min, collecting precipitate, centrifuging and washing with water and anhydrous ethanol for 4 times, drying at 50 ℃ and collecting solid to obtain CeO2And (3) nano materials.
Example 4: bi2S3Preparation of the Dispersion
6mg of flower-like Bi prepared in example 1 were weighed2S3And adding the mixture into 5mL of deionized water, and ultrasonically dispersing for 1 hour.
Example 5: preparation of AgBr Dispersion
6mg of the chitosan functionalized AgBr prepared in example 2 was weighed, added to 6mL of deionized water, and ultrasonically dispersed for 1 hour.
Example 6: CeO (CeO)2Preparation of the Dispersion
6mg of CeO prepared in example 3 were weighed2And adding the mixture into 6mL of deionized water, and ultrasonically dispersing for 1 hour.
Example 7: preparation of electrode washing buffer
10mM Tris-HCl and 50mM KCl were mixed with sterile water as a solvent, and the pH was adjusted to 7.4.
Example 8: preparation of 4-carboxyphenylboronic acid solution
A1 mM 4-carboxyphenylboronic acid solution was prepared using sterile water to prepare a 10mM PBS buffer solution having a pH of 7.4 and a solvent.
Example 9: preparation of EDC, NHS solution
PBS buffer solution with pH 7.4 and concentration 10mM was prepared with sterilized water, and EDC and NHS solutions with concentration 0.5mg/mL were prepared with the PBS solution as solvent, respectively.
Example 10: activation of 4-carboxyphenylboronic acid solution
mu.L of the 4-carboxyphenylboronic acid solution prepared in example 8 was added to 10. mu.L of the EDC and NHS solutions prepared in example 9, respectively, and activated at 37 ℃ for 50min to obtain an activated 4-carboxyphenylboronic acid solution.
Example 11: ITO electrode pretreatment
Cutting ITO conductive glass into 5 × 1cm2With acetone and 1M NaOH in aqueous alcohol (V)Anhydrous ethanolCarrying out ultrasonic treatment on the ITO electrode for 20min by using 1: 1V secondary water and the secondary water, then washing by using the secondary water, and naturally drying for later use.
Example 12: flower-like Bi2S3Is fixed to
40 mu L of Bi2S3And dropwise adding the dispersion liquid to the surface of the pretreated ITO electrode, and irradiating and drying by using an infrared lamp. The electrodes were then washed 3 times with the electrode washing buffer prepared in example 7. Blow-drying with nitrogen. The prepared electrode is marked as Bi2S3/ITO。
Example 13: immobilization of AgBr
Dropwise addition of 40. mu. LAgBr dispersion to Bi2S3Drying the ITO electrode surface by infrared lamp irradiation. Then, the electrode was washed 3 times with the electrode washing buffer prepared in example 7. And (5) drying by nitrogen. The prepared electrode is marked as AgBr/Bi2S3/ITO。
Example 14: immobilization of 4-carboxyphenylboronic acid
20 mu L of activated 4-carboxyphenylboronic acid solution is dripped into AgBr/Bi2S3The ITO electrode surface was reacted with 37 ℃ humidity for 0.5h, and then washed 3 times with the electrode washing buffer prepared in example 7. The prepared electrode is marked as CPBA/AgBr/Bi2S3/ITO。
Example 15: n is a radical of6Immobilization of the antibody to methyladenine
20. mu.L of the suspension was added at a concentration of 25. mu.g.mL-1N of (A)6Dropwise addition of the-methyladenine antibody to CPBA/AgBr/Bi2S3The ITO electrode surface was reacted with 37 ℃ humidity for 1 hour and then washed 3 times with the electrode washing buffer prepared in example 7. The prepared electrode is marked as Ab/CPBA/AgBr/Bi2S3/ITO。
Example 16: n is a radical of6Immobilization of-methyladenine ribonucleotides
20 mu L N6Dropping of-methyladenine ribonucleotides to Ab/CPBA/AgBr/Bi2S3The ITO electrode surface was reacted with 37 ℃ humidity for 1 hour and then washed 3 times with the electrode washing buffer prepared in example 7. The prepared electrode is marked as m6A/m6A/CPBA/AgBr/Bi2S3/ITO。
Example 17: CeO (CeO)2Is fixed to
40 μ L of CeO2Dropping the dispersed liquid to m6A/Ab/CPBA/AgBr/Bi2S3The ITO electrode surface was reacted with 37 ℃ humidity for 2 hours and then washed 3 times with the electrode washing buffer prepared in example 7. The prepared electrode was labeled as CeO2/m6A/m6A/CPBA/AgBr/Bi2S3/ITO。
Example 18: photoelectrochemical detection
An electrochemical workstation is used as a signal acquisition instrument, a 500W xenon lamp is used as a visible light source (a lens for filtering ultraviolet light is additionally arranged), and CeO2/m6A/Ab/CPBA/AgBr/Bi2S3The method comprises the steps of taking an ITO electrode as a working electrode, taking a saturated calomel electrode as a reference electrode, taking a Pt electrode as a counter electrode, taking a PBS (phosphate buffer solution) containing Ascorbic Acid (AA) as an electrode detection solution, taking 0V voltage as a working voltage, adopting an i-t technology to carry out detection research on an object to be detected, and establishing photocurrent intensity and N6Relationship between concentration of methyladenine ribonucleotides (FIG. 2)
Example 19: test for Selectivity
Selectivity is an important indicator of photoelectrochemical sensor performance, and in order to study the specificity of the constructed sensor, the selectivity of the sensor was studied by selecting 5-hydroxycytosine ribonucleotide (5hmC), 5-methylcytosine ribonucleotide (5mC), 5-formylcytosine ribonucleotide (5fC), 5-carboxycytosine ribonucleotide (5caC) and four different bases as interferents. And the change value of the photocurrent (delta I-I) of the sensor constructed by the different interference reagents2-I1，I1Is m6A/Ab/CPBA/AgBr/Bi2S3Current value of/ITO, I2Is m6A/Ab/CPBA/AgBr/Bi2S3The light current values of the electrodes after the ITO was treated with different interferents) were compared. The result shows that the current value change of the sensor constructed by the interferent is obviously lower than N6-methyladenine ribonucleotide, indicating that the constructed sensor has very good specificity (FIG. 3).
Example 20: stability test
The method of the invention is adopted to prepare 7 CeO2/m6A/Ab/CPBA/AgBr/Bi2S3ITO electrode (N)6-methyladenine ribonucleotide concentration of 5-50nM), the photocurrent was detected in the detection solution. The relative standard deviation of the photocurrent obtained was 3.67%, indicating that this method is very goodGood reproducibility. Adding CeO2/m6A/Ab/CPBA/AgBr/Bi2S3The ITO sensor continuously measures for 7 periods, and a photoelectrochemical signal is detected in the detection liquid, so that the standard deviation of the obtained photocurrent is 1.5 percent, which indicates that the method has good stability.
The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims (10)
1. Detect N6-a photoelectrochemical biosensor of methyladenine ribonucleotides, characterised in that it comprises electrodes; the surface of the electrode is sequentially modified with flower-shaped Bi2S3AgBr, 4-carboxyphenylboronic acid, N6-methyladenine antibody, N6-methyladenine ribonucleotides and CeO2。
2. The method for preparing the photoelectrochemical biosensor of claim 1, comprising the steps of:
(1) flower-shaped Bi2S3Fixed on a pretreated electrode to obtain Bi2S3An electrode;
(2) AgBr is fixed on Bi obtained in the step (1)2S3Per electrode surface to obtain AgBr/Bi2S3An electrode;
(3) modifying 4-carboxyl phenylboronic acid on AgBr/Bi obtained in step (2)2S3Per electrode surface to obtain CPBA/AgBr/Bi2S3An electrode;
(4) will N6Modification of the CPBA/AgBr/Bi obtained in step (3) with a methyladenine antibody2S3To obtain Ab/CPBA/AgBr/Bi on the electrode surface2S3An electrode;
(5) using specific binding between antigen and antibody to convert N6-methyladenine ribonucleotide modificationAb/CPBA/AgBr/Bi decorated in the step (4)2S3Per electrode surface, to obtain m6A/Ab/CPBA/AgBr/Bi2S3An electrode;
(6) CeO by utilizing specific combination between phosphate radical and cerium2Modifying m obtained in step (5)6A/Ab/CPBA/AgBr/Bi2S3On the surface of an electrode to obtain CeO2/m6A/Ab/CPBA/AgBr/Bi2S3Electrode, i.e. detecting N6-a photoelectrochemical biosensor of methyladenine ribonucleotides.
3. The method according to claim 2, wherein in the step (1), Bi is flower-shaped2S3The fixing method comprises the following steps:
flower-shaped Bi2S3The nano material is evenly dispersed in deionized water to obtain Bi2S3A dispersion of Bi2S3Dripping the dispersed liquid on the surface of a pretreatment electrode, and drying under the irradiation of an infrared lamp;
preferably, said flower-like Bi2S3The nano material is prepared by the following method:
separately adding Bi (NO)3)3·5H2Dissolving O, thiourea, urea and 4-carboxyphenylboronic acid in ethylene glycol, carrying out hydrothermal reaction, collecting precipitate, centrifugally washing with water and ethanol, drying, and collecting solid to obtain flower-shaped Bi2S3And (3) nano materials.
4. The preparation method according to claim 2, wherein in the step (2), AgBr is fixed by:
uniformly dispersing AgBr nano material in deionized water to obtain AgBr dispersion liquid, and dropwise adding the AgBr dispersion liquid into the Bi obtained in the step (1)2S3Drying the electrode surface under the irradiation of an infrared lamp;
preferably, the AgBr nano material is prepared by the following method:
mixing AgNO3Mixing the aqueous solution and the chitosan aqueous solution, adding the KBr aqueous solution in the stirring process, and stirring for 1-And 5h later, forming a dispersion containing AgBr nano-particles, centrifugally collecting precipitates, centrifugally washing with water and ethanol, drying and collecting solids to obtain the AgBr nano-material.
5. The method according to claim 2, wherein in the step (3), the 4-carboxyphenylboronic acid is modified by:
dropwise adding the activated 4-carboxyphenylboronic acid solution to AgBr/Bi obtained in the step (2)2S3The surface of the electrode is reacted for 0.5 to 3 hours under the humid condition of 37 ℃ and then is cleaned for 1 to 5 times;
preferably, the activated 4-carboxyphenylboronic acid solution is prepared by the following method: respectively adding 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and an N-hydroxysuccinimide solution into the 4-carboxyphenylboronic acid solution, and activating at 37 ℃ for 10-90min to obtain an activated 4-carboxyphenylboronic acid solution; the 4-carboxyphenylboronic acid solution is prepared by PBS buffer solution with pH of 5.5-8.5 and concentration of 5-500mM, and the concentration of the 4-carboxyphenylboronic acid solution is 0.5-5 mM; 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide solution are respectively prepared by PBS buffer solution with pH of 5.5-8.5 and concentration of 5-500mM, and the concentrations of the 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and the N-hydroxysuccinimide solution are 0.5mg/mL-1。
6. The method according to claim 2, wherein in the step (4), N is6The method for modifying the-methyladenine antibody comprises the following steps:
will N6Dropwise adding a methyl adenine antibody solution to the CPBA/AgBr/Bi obtained in the step (3)2S3The surface of the electrode is reacted for 0.5 to 3 hours under the humid condition of 37 ℃ and then washed for 1 to 5 times.
7. The method according to claim 2, wherein in the step (5), N is6The method for modifying the-methyladenine ribonucleotide comprises the following steps:
will N6-dropwise addition of a solution of methyladenine ribonucleotides to the stepAb/CPBA/AgBr/Bi obtained in step (4)2S3The surface of the electrode is reacted for 0.5 to 3 hours under the humid condition of 37 ℃ and then washed for 1 to 5 times.
8. The production method according to claim 2, wherein, in the step (6), CeO is used2The modification method comprises the following steps:
adding CeO2Dropwise adding the nano material dispersion liquid to the m obtained in the step (5)6A/Ab/CPBA/AgBr/Bi2S3The surface of the electrode is reacted for 0.5 to 5 hours under the humid condition of 37 ℃ and then is cleaned for 1 to 5 times;
preferably, the CeO2The nano material is prepared by the following method:
respectively adding Na3PO4·12H2O and Ce (NO)3)3·6H2Dissolving O in deionized water, carrying out hydrothermal reaction, collecting precipitate, centrifugally washing with water and ethanol, drying, and collecting solid to obtain CeO2And (3) nano materials.
9. The photoelectrochemical biosensor of claim 1 detecting N6-methyladenine ribonucleotides.
10. Detection of N using the photoelectrochemical biosensor of claim 16-a method of producing methyladenine ribonucleotides, characterized in that it comprises the following steps:
CeO with photoelectrochemical biosensor2/m6A/Ab/CPBA/AgBr/Bi2S3The electrode is used as a working electrode, the saturated calomel electrode is used as a reference electrode, the Pt wire is used as an auxiliary electrode to form a three-electrode system for photoelectric chemical signal detection, the electrode detection solution is a PBS (phosphate buffer solution) containing ascorbic acid, and current and N are established6Relationship between concentrations of methyladenine ribonucleotides for N6-detecting the content of methyladenine ribonucleotides;
preferably, the PBS buffer solution has a pH of 5.5-8.5 and a concentration of 5-500 mM;
preferably, for N6-methyl glandThe method for detecting the content of purine ribonucleotide is a current-time method, and the applied potential is as follows: -0.5-0.5V.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113740401A (en) * | 2021-07-07 | 2021-12-03 | 中国人民解放军陆军军医大学 | M in RNA5C and m6A double analysis logic photoelectrochemical sensing method |
| CN114137053A (en) * | 2021-11-30 | 2022-03-04 | 山东农业大学 | Antibody enzyme-free assisted photoelectrochemical sensor for detecting m6Method A |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100180949A1 (en) * | 2005-09-22 | 2010-07-22 | Sumitomo Chemical Company, Limited | Compound, photoelectric converter and photoelectrochemical cell |
| CN108918622A (en) * | 2018-05-18 | 2018-11-30 | 山东农业大学 | A kind of Photoelectrochemistrbiosensor biosensor and preparation method thereof detecting 5-hydroxymethyl cytosine deoxyribonucleotide |
| CN109709170A (en) * | 2019-01-18 | 2019-05-03 | 山东农业大学 | A method of the photoelectrochemical assay based on black titanium dioxide detects 5-hydroxymethyl cytosine |
| CN109828006A (en) * | 2019-02-27 | 2019-05-31 | 山东农业大学 | A kind of the optical electro-chemistry sensor and its detection method of detection methylation RNA |
| CN110426428A (en) * | 2019-07-30 | 2019-11-08 | 山东农业大学 | Optical electro-chemistry sensor based on Ti-MOF detects N1The method of methyl adenine |
-
2021
- 2021-02-24 CN CN202110205043.3A patent/CN112858417B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100180949A1 (en) * | 2005-09-22 | 2010-07-22 | Sumitomo Chemical Company, Limited | Compound, photoelectric converter and photoelectrochemical cell |
| CN108918622A (en) * | 2018-05-18 | 2018-11-30 | 山东农业大学 | A kind of Photoelectrochemistrbiosensor biosensor and preparation method thereof detecting 5-hydroxymethyl cytosine deoxyribonucleotide |
| CN109709170A (en) * | 2019-01-18 | 2019-05-03 | 山东农业大学 | A method of the photoelectrochemical assay based on black titanium dioxide detects 5-hydroxymethyl cytosine |
| CN109828006A (en) * | 2019-02-27 | 2019-05-31 | 山东农业大学 | A kind of the optical electro-chemistry sensor and its detection method of detection methylation RNA |
| CN110426428A (en) * | 2019-07-30 | 2019-11-08 | 山东农业大学 | Optical electro-chemistry sensor based on Ti-MOF detects N1The method of methyl adenine |
Non-Patent Citations (2)
| Title |
|---|
| YUE WANG等: "Photoelectrochemical immunosensor for N6-methyladenine detection based on Ru@UiO-66, Bi2O3 and Black TiO2", 《BIOSENSORS AND BIOELECTRONICS》 * |
| YUNLEI ZHOU等: "A novel photoelectrochemical immunosensor for N1-methyladenine detection based on BiVO4/g-C3N4 heterojunction with signal amplification of TiO2@NH2-MIL-125(Ti)", 《SENSORS AND ACTUATORS B: CHEMICAL》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113740401A (en) * | 2021-07-07 | 2021-12-03 | 中国人民解放军陆军军医大学 | M in RNA5C and m6A double analysis logic photoelectrochemical sensing method |
| CN113740401B (en) * | 2021-07-07 | 2024-03-29 | 中国人民解放军陆军军医大学 | M in RNA 5 C and m 6 A dual analysis logic photoelectrochemical sensing method |
| CN114137053A (en) * | 2021-11-30 | 2022-03-04 | 山东农业大学 | Antibody enzyme-free assisted photoelectrochemical sensor for detecting m6Method A |
| CN114137053B (en) * | 2021-11-30 | 2022-08-19 | 山东农业大学 | Antibody enzyme-free assisted photoelectrochemical sensor for detecting m 6 Method A |
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